To determine whether major differences in vascular endothelial growth factor secretion exist betw... more To determine whether major differences in vascular endothelial growth factor secretion exist between adenocarcinomas of the uterine cervix compared with squamous cell carcinomas. The secretion of vascular endothelial growth factor by eight fresh cervical cancer cell preparations (four adenocarcinomas and four squamous cell carcinomas) and four established squamous cell lines was evaluated using a sensitive enzyme-linked immunosorbent assay in vitro. All cervical tumors secreted significant amounts of vascular endothelial growth factor, and no significant differences between fresh and established squamous cell lines were detectable. In contrast, a highly significant difference in vascular endothelial growth factor secretion was noted between fresh adenocarcinomas (mean = 2712, range between 1700 to 3500 pg/mL/10(5) cells/48 hours) when compared with fresh squamous (mean = 575, range between 200 to 950 pg/mL/10(5) cells/48 hours) or established squamous cervical carcinoma cell lines (mean = 712, range between 400 to 1000 pg/mL/10(5) cells/48 hours) (F-test, P< or =.001). These data strongly suggest that major differences in the secretion of vascular endothelial growth factor exist between squamous cell carcinoma and adenocarcinomas of the uterine cervix. Therefore, at least some of the differences in the natural biologic behavior of these two histologic types of cervical cancer, including the propensity for earlier lymphatic and hematogenous metastasis as well as the lower response to radiation treatment, could be related to major differences in the secretion of this powerful angiogenic and immunosuppressive cytokine.
Objective: To determine whether major differences in vascular endothelial growth factor secretion... more Objective: To determine whether major differences in vascular endothelial growth factor secretion exist between adenocarcinomas of the uterine cervix compared with squamous cell carcinomas.Methods: The secretion of vascular endothelial growth factor by eight fresh cervical cancer cell preparations (four adenocarcinomas and four squamous cell carcinomas) and four established squamous cell lines was evaluated using a sensitive enzyme-linked immunosorbent assay in
The aim of this study was to compare the phenotype and function of lymphocytes collected from the... more The aim of this study was to compare the phenotype and function of lymphocytes collected from the peripheral blood (PBL), tumor draining regional lymph nodes (LND), and infiltrating tumor tissues (TIL) of patients with stage IB-IIA cervical cancer. Leukocytes from peripheral blood (n = 35), tumor draining lymph nodes (n = 33), and tumor tissues (n = 15) of cervical cancer patients were evaluated for the relative proportions of lymphocyte subsets including CD3+, CD4+, CD8+, CD19+, CD56, and the early (CD25) and late (HLA-DR) activation markers on CD3+ T cells, as well as the ability to synthesize type 1 cytokines (IFN-gamma and IL-2) and a type 2 cytokine (IL-4) by flow cytometry. In all patients, T cells (CD3+) were the major leukocyte population detected in each tissue, with CD4+ T cells being dominant in PBL and LND, while CD8+ T cells predominated in TIL (CD4:CD8 ratios, 2.4 vs 4.0 vs 0.7, respectively). CD19+ lymphocytes (B cells) were significantly higher in LND compared to PBL and TIL (P > 0.01) while CD56+ lymphocytes were higher in PBL compared to LND (P > 0.01) and TIL (P > 0.05). The early activation marker CD25 was significantly up-regulated in LND, while TIL had a higher proportion of T cells expressing the late activation marker HLA-DR. Type 1 cytokines were the dominant type produced by in vitro stimulated T cells for each population, with a greater proportion of IFN-gamma+ CD4+ and CD8+ T cells (i.e., Th1 and Tc1) and IL-2+ CD8+ T cells (Tc1) seen in TIL, as compared with LND and PBL (P > 0.01). Low percentages of IL-4+ T cells (i.e., Th2 and Tc2) were detected only in PBL. This study demonstrates significant differences in the phenotype and activation state of lymphocyte subsets from different anatomical sites, as well as differences in their ability to synthesize immunostimulatory cytokines. The recruitment and accumulation of high concentrations of antigen-experienced T lymphocytes in the cervical tumor tissue may represent an important local barrier to neoplastic dissemination.
Seventy-five women with anti-hepatitis C virus (HCV) antibody were enrolled prospectively during ... more Seventy-five women with anti-hepatitis C virus (HCV) antibody were enrolled prospectively during pregnancy or at delivery for study of mother-to-child transmission of HCV. Twenty-three women were coinfected with the human immunodeficiency virus (HIV). Seventy babies were monitored for at least 6 months. HCV infection was diagnosed in six infants (8.6%), four of whom were born to anti-HIV-positive mothers. HCV RNA was first detected between 2 and 6 months, and the genotypes of infected babies matched those of their mothers (type 1: n = 4; type 3: n = 2). Identical master sequences of the hypervariable region (HVR1) were detected in a mother-infant pair. In three babies coinfected with HCV and HIV, anti-HCV disappeared between 2 and 7 months, being persistently negative in two cases monitored for 11 and 26 months. Transmitting mothers did not differ significantly from those who did not transmit the infection with anti-HIV, HCV genotypes, and viral load at delivery, but had lower rate of reactivity to C100 by the recombinant immunoblot assay (RIBA) (P < .01). This prospective study confirms transmission of HCV from anti-HIV-negative mothers (4.4% in this series). Absence of anti-C100 antibodies at delivery is apparently related to increased risk of vertical transmission. Seronegative HCV infection can be observed in children coinfected with HIV.
We have evaluated the clinical significance of hepatitis C virus (HCV) RNA determination by analy... more We have evaluated the clinical significance of hepatitis C virus (HCV) RNA determination by analyzing a group of 221 hospitalized patients with abnormal liver function tests. Serum HCV RNA was detected by "nested" PCR amplification followed by nonisotopic hybridization. Of the 200 (90.5%) patients with anti-HCV-positive enzyme-linked immunosorbent assay results, 152 (76%) were RIBA reactive, 47 (23.5%) had indeterminate results, and 1 (0.5%) was nonreactive. Of the 180 (90%) patients positive for anti-HCV and HCV RNA, 138 (76.7%) were RIBA reactive and 42 (23.3%) were RIBA indeterminate. The pattern of RIBA reactivity did not correlate with the presence of HCV RNA. Elevated alanine aminotransferase levels were associated neither with the presence of viremia nor with the RIBA pattern. Histological findings consistent with non-A non-B hepatitis correlated with the presence of HCV RNA but not with the RIBA pattern. HCV RNA was detected in 11 of 21 (52.4%) anti-HCV-negative pa...
Introduction/Background High-grade serous ovarian carcinoma (HGSOC), the most common and deadly e... more Introduction/Background High-grade serous ovarian carcinoma (HGSOC), the most common and deadly epithelial ovarian cancer histotype is characterized by a heterogeneous genomic landscape. After surgery, all patients are treated with adjuvant platinum (Pt)-based chemotherapy to which the response is heterogeneous, ranging from cases sensitive (Platinum-sensitive, Pt-s cases), to intrinsically resistant patients, who relapse within 6 months from the end of therapy (Pt-resistant, Pt-r). The aim of the study was to investigate the mechanisms characterizing the biology of primary resistance to upfront Pt-based chemotherapy through an integrated pathway analysis. Methodology Gene and miRNA microarray experiments were carried out on 36 Pt-s and 41 Pt-r tumor samples. Gene and miRNA expression have been integrated using Micrographite algorithm. Expression levels of selected predictive and prognostic genes were validated on a total of 242 HGSOC samples by qRT-PCR and on the Curated Ovarian Database (including 838 HGSOC). Results Expression profiles of 131 mRNAs and five miRNAs, belonging to five different and functionally-related molecular pathways, discriminate Pt-s and Pt-r cases. We selected 23 elements of the networks for orthogonal validation and 19 coding genes confirmed as differentially expressed between Pt-r and Pt-s cases. Among them, 16 elements also showed independent prognostic value in terms of PFS and OS in multivariate analysis. The prognostic impact of this signature was in silico validated on the Curated Ovarian Database, resulting in a 3-gene signature as independent prognostic biomarker of survival for HGSOC. Conclusion In the current study, we used an integrated pathway analysis on miRNA and gene expression signatures to capture the key pathways shaping the different biology of Pt-r and Pt-s tumors, validating our results using two independent cohorts of HGSOC biopsies. Finally, we investigated the association of the validated signature with survival across multiple HGSOC databases. This strategy identified a three-gene signature with predictive and prognostic impact in HGSOC. Disclosure Nothing to disclose
For high‐risk endometrial cancer (EC) patients, adjuvant chemotherapy is recommended to improve o... more For high‐risk endometrial cancer (EC) patients, adjuvant chemotherapy is recommended to improve outcome. Yet, predictive biomarkers for response to platinum‐based chemotherapy (Pt‐aCT) are currently lacking. We tested expression of L1 cell‐adhesion molecule (L1CAM), a well‐recognised marker of poor prognosis in EC, in tumour samples from high‐risk EC patients, to explore its role as a predictive marker of Pt‐aCT response. L1CAM expression was determined using RT‐qPCR and immunohistochemistry in a cohort of high‐risk EC patients treated with Pt‐aCT and validated in a multicentric independent cohort. The association between L1CAM and clinicopathologic features and L1CAM additive value in predicting platinum response were determined. The effect of L1CAM gene silencing on response to carboplatin was functionally tested on primary L1CAM‐expressing cells. Increased L1CAM expression at both genetic and protein level correlated with high‐grade, non‐endometrioid histology and poor response to platinum treatment. A predictive model adding L1CAM to prognostic clinical variables significantly improved platinum response prediction (C‐index 78.1%, P = .012). In multivariate survival analysis, L1CAM expression was significantly associated with poor outcome (HR: 2.03, P = .019), potentially through an indirect effect, mediated by its influence on response to chemotherapy. In vitro, inhibition of L1CAM significantly increased cell sensitivity to carboplatin, supporting a mechanistic link between L1CAM expression and response to platinum in EC cells. In conclusion, we have demonstrated the role of L1CAM in the prediction of response to Pt‐aCT in two independent cohorts of high‐risk EC patients. L1CAM is a promising candidate biomarker to optimise decision making in high‐risk patients who are eligible for Pt‐aCT.
To identify novel molecular biomarkers useful for the early diagnosis and therapy of ovarian canc... more To identify novel molecular biomarkers useful for the early diagnosis and therapy of ovarian cancer by gene expression profiling. To compare the genetic fingerprints of flash-frozen ovarian serous carcinomas to those of matched highly purified primary tumor cell cultures. Gene expression profiles of 19 flash-frozen ovarian serous papillary carcinoma (OSPC) were analyzed and compared to 15 controls (highly purified human ovarian surface epithelium short-term cultures, HOSE) using oligonucleotide microarrays complementary to &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;14,500 human genes. In addition, gene expression profiling of 5 highly purified primary OSPC cultured in vitro for less than 2 weeks was compared to flash-frozen ovarian carcinoma biopsies obtained from matched samples. Quantitative RT-PCR and IHC staining techniques were used to validate microarray data at RNA and protein levels for some of the differentially expressed genes. Unsupervised analysis of gene expression data readily distinguished normal tissue from flash-frozen OSPC and identified 901 and 557 genes that exhibited &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;3-fold up-regulation or down-regulation, respectively, in OSPC when compared to HOSE. Mammaglobin 2, an ovarian secreted protein, was identified as the top differentially expressed gene in OSPC (19 out 19 OSPC versus 0 out of 15 HOSE) with over 827-fold up-regulation relative to HOSE. The claudin and kallikrein family of proteins including the clostridium perfringens enterotoxin receptors claudin 3 and 4, kallikreins 6, 7, 8, 10, 11 and the immunomodulatory molecule B7-H4 were found among the most highly overexpressed genes in OSPC when compared to HOSE. Genetic fingerprints of flash-frozen OSPC were found to have high correlation with those of purified primary OSPC short-term in vitro cultures with only 31 out of 8,637 genes (0.35%) differentially expressed between the two groups. Short-term in vitro culture of primary ovarian carcinomas may greatly increase the purity of ovarian tumor RNA available for gene expression profiling without causing major alteration in OSPC fingerprints. Mammaglobin 2, kallikreins 6, 7, 8, 10, 11, claudin 3 and 4 and B7-H4 gene expression products represent candidate biomarkers endowed with great potential for early screening and therapy of OSPC patients.
Supplementary data, material and results. Figure S1.1. Overall analysis schema Table S1.1 REMARK ... more Supplementary data, material and results. Figure S1.1. Overall analysis schema Table S1.1 REMARK form. Figure S1.2 a) Kaplan Meier curves of patients divided by tumor grades (OS and PFS model). The table below summarizes the models used. Figure S1.2 b) Kaplan Meier curves of patients divided in patients who received chemotherapy (Yes) and patients who did not undergo chemotherapy (No). Figure S1.2 c) Kaplan Meier curves of patients divided in FIGO substages. The table below summarizes the models used.
PDF file - 217K, The clinical stratification of the 76 selected patients used for the integrative... more PDF file - 217K, The clinical stratification of the 76 selected patients used for the integrative analysis and the MAGIA2 complete results.
To determine whether major differences in vascular endothelial growth factor secretion exist betw... more To determine whether major differences in vascular endothelial growth factor secretion exist between adenocarcinomas of the uterine cervix compared with squamous cell carcinomas. The secretion of vascular endothelial growth factor by eight fresh cervical cancer cell preparations (four adenocarcinomas and four squamous cell carcinomas) and four established squamous cell lines was evaluated using a sensitive enzyme-linked immunosorbent assay in vitro. All cervical tumors secreted significant amounts of vascular endothelial growth factor, and no significant differences between fresh and established squamous cell lines were detectable. In contrast, a highly significant difference in vascular endothelial growth factor secretion was noted between fresh adenocarcinomas (mean = 2712, range between 1700 to 3500 pg/mL/10(5) cells/48 hours) when compared with fresh squamous (mean = 575, range between 200 to 950 pg/mL/10(5) cells/48 hours) or established squamous cervical carcinoma cell lines (mean = 712, range between 400 to 1000 pg/mL/10(5) cells/48 hours) (F-test, P&amp;amp;amp;amp;amp;amp;amp;amp;lt; or =.001). These data strongly suggest that major differences in the secretion of vascular endothelial growth factor exist between squamous cell carcinoma and adenocarcinomas of the uterine cervix. Therefore, at least some of the differences in the natural biologic behavior of these two histologic types of cervical cancer, including the propensity for earlier lymphatic and hematogenous metastasis as well as the lower response to radiation treatment, could be related to major differences in the secretion of this powerful angiogenic and immunosuppressive cytokine.
Objective: To determine whether major differences in vascular endothelial growth factor secretion... more Objective: To determine whether major differences in vascular endothelial growth factor secretion exist between adenocarcinomas of the uterine cervix compared with squamous cell carcinomas.Methods: The secretion of vascular endothelial growth factor by eight fresh cervical cancer cell preparations (four adenocarcinomas and four squamous cell carcinomas) and four established squamous cell lines was evaluated using a sensitive enzyme-linked immunosorbent assay in
The aim of this study was to compare the phenotype and function of lymphocytes collected from the... more The aim of this study was to compare the phenotype and function of lymphocytes collected from the peripheral blood (PBL), tumor draining regional lymph nodes (LND), and infiltrating tumor tissues (TIL) of patients with stage IB-IIA cervical cancer. Leukocytes from peripheral blood (n = 35), tumor draining lymph nodes (n = 33), and tumor tissues (n = 15) of cervical cancer patients were evaluated for the relative proportions of lymphocyte subsets including CD3+, CD4+, CD8+, CD19+, CD56, and the early (CD25) and late (HLA-DR) activation markers on CD3+ T cells, as well as the ability to synthesize type 1 cytokines (IFN-gamma and IL-2) and a type 2 cytokine (IL-4) by flow cytometry. In all patients, T cells (CD3+) were the major leukocyte population detected in each tissue, with CD4+ T cells being dominant in PBL and LND, while CD8+ T cells predominated in TIL (CD4:CD8 ratios, 2.4 vs 4.0 vs 0.7, respectively). CD19+ lymphocytes (B cells) were significantly higher in LND compared to PBL and TIL (P &gt; 0.01) while CD56+ lymphocytes were higher in PBL compared to LND (P &gt; 0.01) and TIL (P &gt; 0.05). The early activation marker CD25 was significantly up-regulated in LND, while TIL had a higher proportion of T cells expressing the late activation marker HLA-DR. Type 1 cytokines were the dominant type produced by in vitro stimulated T cells for each population, with a greater proportion of IFN-gamma+ CD4+ and CD8+ T cells (i.e., Th1 and Tc1) and IL-2+ CD8+ T cells (Tc1) seen in TIL, as compared with LND and PBL (P &gt; 0.01). Low percentages of IL-4+ T cells (i.e., Th2 and Tc2) were detected only in PBL. This study demonstrates significant differences in the phenotype and activation state of lymphocyte subsets from different anatomical sites, as well as differences in their ability to synthesize immunostimulatory cytokines. The recruitment and accumulation of high concentrations of antigen-experienced T lymphocytes in the cervical tumor tissue may represent an important local barrier to neoplastic dissemination.
Seventy-five women with anti-hepatitis C virus (HCV) antibody were enrolled prospectively during ... more Seventy-five women with anti-hepatitis C virus (HCV) antibody were enrolled prospectively during pregnancy or at delivery for study of mother-to-child transmission of HCV. Twenty-three women were coinfected with the human immunodeficiency virus (HIV). Seventy babies were monitored for at least 6 months. HCV infection was diagnosed in six infants (8.6%), four of whom were born to anti-HIV-positive mothers. HCV RNA was first detected between 2 and 6 months, and the genotypes of infected babies matched those of their mothers (type 1: n = 4; type 3: n = 2). Identical master sequences of the hypervariable region (HVR1) were detected in a mother-infant pair. In three babies coinfected with HCV and HIV, anti-HCV disappeared between 2 and 7 months, being persistently negative in two cases monitored for 11 and 26 months. Transmitting mothers did not differ significantly from those who did not transmit the infection with anti-HIV, HCV genotypes, and viral load at delivery, but had lower rate of reactivity to C100 by the recombinant immunoblot assay (RIBA) (P < .01). This prospective study confirms transmission of HCV from anti-HIV-negative mothers (4.4% in this series). Absence of anti-C100 antibodies at delivery is apparently related to increased risk of vertical transmission. Seronegative HCV infection can be observed in children coinfected with HIV.
We have evaluated the clinical significance of hepatitis C virus (HCV) RNA determination by analy... more We have evaluated the clinical significance of hepatitis C virus (HCV) RNA determination by analyzing a group of 221 hospitalized patients with abnormal liver function tests. Serum HCV RNA was detected by "nested" PCR amplification followed by nonisotopic hybridization. Of the 200 (90.5%) patients with anti-HCV-positive enzyme-linked immunosorbent assay results, 152 (76%) were RIBA reactive, 47 (23.5%) had indeterminate results, and 1 (0.5%) was nonreactive. Of the 180 (90%) patients positive for anti-HCV and HCV RNA, 138 (76.7%) were RIBA reactive and 42 (23.3%) were RIBA indeterminate. The pattern of RIBA reactivity did not correlate with the presence of HCV RNA. Elevated alanine aminotransferase levels were associated neither with the presence of viremia nor with the RIBA pattern. Histological findings consistent with non-A non-B hepatitis correlated with the presence of HCV RNA but not with the RIBA pattern. HCV RNA was detected in 11 of 21 (52.4%) anti-HCV-negative pa...
Introduction/Background High-grade serous ovarian carcinoma (HGSOC), the most common and deadly e... more Introduction/Background High-grade serous ovarian carcinoma (HGSOC), the most common and deadly epithelial ovarian cancer histotype is characterized by a heterogeneous genomic landscape. After surgery, all patients are treated with adjuvant platinum (Pt)-based chemotherapy to which the response is heterogeneous, ranging from cases sensitive (Platinum-sensitive, Pt-s cases), to intrinsically resistant patients, who relapse within 6 months from the end of therapy (Pt-resistant, Pt-r). The aim of the study was to investigate the mechanisms characterizing the biology of primary resistance to upfront Pt-based chemotherapy through an integrated pathway analysis. Methodology Gene and miRNA microarray experiments were carried out on 36 Pt-s and 41 Pt-r tumor samples. Gene and miRNA expression have been integrated using Micrographite algorithm. Expression levels of selected predictive and prognostic genes were validated on a total of 242 HGSOC samples by qRT-PCR and on the Curated Ovarian Database (including 838 HGSOC). Results Expression profiles of 131 mRNAs and five miRNAs, belonging to five different and functionally-related molecular pathways, discriminate Pt-s and Pt-r cases. We selected 23 elements of the networks for orthogonal validation and 19 coding genes confirmed as differentially expressed between Pt-r and Pt-s cases. Among them, 16 elements also showed independent prognostic value in terms of PFS and OS in multivariate analysis. The prognostic impact of this signature was in silico validated on the Curated Ovarian Database, resulting in a 3-gene signature as independent prognostic biomarker of survival for HGSOC. Conclusion In the current study, we used an integrated pathway analysis on miRNA and gene expression signatures to capture the key pathways shaping the different biology of Pt-r and Pt-s tumors, validating our results using two independent cohorts of HGSOC biopsies. Finally, we investigated the association of the validated signature with survival across multiple HGSOC databases. This strategy identified a three-gene signature with predictive and prognostic impact in HGSOC. Disclosure Nothing to disclose
For high‐risk endometrial cancer (EC) patients, adjuvant chemotherapy is recommended to improve o... more For high‐risk endometrial cancer (EC) patients, adjuvant chemotherapy is recommended to improve outcome. Yet, predictive biomarkers for response to platinum‐based chemotherapy (Pt‐aCT) are currently lacking. We tested expression of L1 cell‐adhesion molecule (L1CAM), a well‐recognised marker of poor prognosis in EC, in tumour samples from high‐risk EC patients, to explore its role as a predictive marker of Pt‐aCT response. L1CAM expression was determined using RT‐qPCR and immunohistochemistry in a cohort of high‐risk EC patients treated with Pt‐aCT and validated in a multicentric independent cohort. The association between L1CAM and clinicopathologic features and L1CAM additive value in predicting platinum response were determined. The effect of L1CAM gene silencing on response to carboplatin was functionally tested on primary L1CAM‐expressing cells. Increased L1CAM expression at both genetic and protein level correlated with high‐grade, non‐endometrioid histology and poor response to platinum treatment. A predictive model adding L1CAM to prognostic clinical variables significantly improved platinum response prediction (C‐index 78.1%, P = .012). In multivariate survival analysis, L1CAM expression was significantly associated with poor outcome (HR: 2.03, P = .019), potentially through an indirect effect, mediated by its influence on response to chemotherapy. In vitro, inhibition of L1CAM significantly increased cell sensitivity to carboplatin, supporting a mechanistic link between L1CAM expression and response to platinum in EC cells. In conclusion, we have demonstrated the role of L1CAM in the prediction of response to Pt‐aCT in two independent cohorts of high‐risk EC patients. L1CAM is a promising candidate biomarker to optimise decision making in high‐risk patients who are eligible for Pt‐aCT.
To identify novel molecular biomarkers useful for the early diagnosis and therapy of ovarian canc... more To identify novel molecular biomarkers useful for the early diagnosis and therapy of ovarian cancer by gene expression profiling. To compare the genetic fingerprints of flash-frozen ovarian serous carcinomas to those of matched highly purified primary tumor cell cultures. Gene expression profiles of 19 flash-frozen ovarian serous papillary carcinoma (OSPC) were analyzed and compared to 15 controls (highly purified human ovarian surface epithelium short-term cultures, HOSE) using oligonucleotide microarrays complementary to &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;14,500 human genes. In addition, gene expression profiling of 5 highly purified primary OSPC cultured in vitro for less than 2 weeks was compared to flash-frozen ovarian carcinoma biopsies obtained from matched samples. Quantitative RT-PCR and IHC staining techniques were used to validate microarray data at RNA and protein levels for some of the differentially expressed genes. Unsupervised analysis of gene expression data readily distinguished normal tissue from flash-frozen OSPC and identified 901 and 557 genes that exhibited &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;3-fold up-regulation or down-regulation, respectively, in OSPC when compared to HOSE. Mammaglobin 2, an ovarian secreted protein, was identified as the top differentially expressed gene in OSPC (19 out 19 OSPC versus 0 out of 15 HOSE) with over 827-fold up-regulation relative to HOSE. The claudin and kallikrein family of proteins including the clostridium perfringens enterotoxin receptors claudin 3 and 4, kallikreins 6, 7, 8, 10, 11 and the immunomodulatory molecule B7-H4 were found among the most highly overexpressed genes in OSPC when compared to HOSE. Genetic fingerprints of flash-frozen OSPC were found to have high correlation with those of purified primary OSPC short-term in vitro cultures with only 31 out of 8,637 genes (0.35%) differentially expressed between the two groups. Short-term in vitro culture of primary ovarian carcinomas may greatly increase the purity of ovarian tumor RNA available for gene expression profiling without causing major alteration in OSPC fingerprints. Mammaglobin 2, kallikreins 6, 7, 8, 10, 11, claudin 3 and 4 and B7-H4 gene expression products represent candidate biomarkers endowed with great potential for early screening and therapy of OSPC patients.
Supplementary data, material and results. Figure S1.1. Overall analysis schema Table S1.1 REMARK ... more Supplementary data, material and results. Figure S1.1. Overall analysis schema Table S1.1 REMARK form. Figure S1.2 a) Kaplan Meier curves of patients divided by tumor grades (OS and PFS model). The table below summarizes the models used. Figure S1.2 b) Kaplan Meier curves of patients divided in patients who received chemotherapy (Yes) and patients who did not undergo chemotherapy (No). Figure S1.2 c) Kaplan Meier curves of patients divided in FIGO substages. The table below summarizes the models used.
PDF file - 217K, The clinical stratification of the 76 selected patients used for the integrative... more PDF file - 217K, The clinical stratification of the 76 selected patients used for the integrative analysis and the MAGIA2 complete results.
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