CN108018354A - MicroRNA-34在抑制前列腺癌转移治疗中的新用途 - Google Patents
MicroRNA-34在抑制前列腺癌转移治疗中的新用途 Download PDFInfo
- Publication number
- CN108018354A CN108018354A CN201711370384.6A CN201711370384A CN108018354A CN 108018354 A CN108018354 A CN 108018354A CN 201711370384 A CN201711370384 A CN 201711370384A CN 108018354 A CN108018354 A CN 108018354A
- Authority
- CN
- China
- Prior art keywords
- mir
- prostate cancer
- cells
- microrna
- invasion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 206010060862 Prostate cancer Diseases 0.000 title description 13
- 208000000236 Prostatic Neoplasms Diseases 0.000 title description 13
- 108091079013 miR-34b Proteins 0.000 claims abstract description 27
- 108091084018 miR-34b stem-loop Proteins 0.000 claims abstract description 27
- 108091063470 miR-34b-1 stem-loop Proteins 0.000 claims abstract description 27
- 108091049916 miR-34b-2 stem-loop Proteins 0.000 claims abstract description 27
- 108091057222 miR-34b-3 stem-loop Proteins 0.000 claims abstract description 27
- 108091092639 miR-34b-4 stem-loop Proteins 0.000 claims abstract description 27
- 108091090583 miR-34c stem-loop Proteins 0.000 claims abstract description 16
- 108091082133 miR-34c-1 stem-loop Proteins 0.000 claims abstract description 16
- 208000010658 metastatic prostate carcinoma Diseases 0.000 claims abstract description 14
- 239000000090 biomarker Substances 0.000 claims abstract description 5
- 230000005012 migration Effects 0.000 claims description 14
- 238000013508 migration Methods 0.000 claims description 14
- 230000009545 invasion Effects 0.000 claims description 13
- 239000003112 inhibitor Substances 0.000 claims description 11
- 206010027476 Metastases Diseases 0.000 claims description 8
- 230000009401 metastasis Effects 0.000 claims description 8
- 206010028980 Neoplasm Diseases 0.000 claims description 6
- 201000011510 cancer Diseases 0.000 claims description 6
- 230000005764 inhibitory process Effects 0.000 claims description 3
- 230000004709 cell invasion Effects 0.000 abstract description 4
- 230000012292 cell migration Effects 0.000 abstract description 4
- 230000003278 mimic effect Effects 0.000 description 8
- 230000014509 gene expression Effects 0.000 description 7
- 239000013642 negative control Substances 0.000 description 7
- 239000002609 medium Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 238000001890 transfection Methods 0.000 description 5
- 239000012124 Opti-MEM Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 230000001394 metastastic effect Effects 0.000 description 3
- 206010061289 metastatic neoplasm Diseases 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108700011259 MicroRNAs Proteins 0.000 description 2
- 108010082117 matrigel Proteins 0.000 description 2
- 238000010232 migration assay Methods 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 108091074487 miR-34 stem-loop Proteins 0.000 description 1
- 108091092493 miR-34-1 stem-loop Proteins 0.000 description 1
- 108091059780 miR-34-2 stem-loop Proteins 0.000 description 1
- 108091029119 miR-34a stem-loop Proteins 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Pathology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Oncology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Hospice & Palliative Care (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明公开了miR‑34b和miR‑34c抑制转移性前列腺癌细胞迁移和侵袭。为治疗转移性前列腺癌提供新的生物学标志物。
Description
技术领域
本发明涉及细胞生物学、基因组学及生物信息学领域的相关技术。
背景技术
MicroRNA（miRNA）作为重要的基因表达调控分子，在肿瘤发生过程中可以发挥癌基因或抑癌基因的作用。miR-34家族有三个成员分别是miR-34a、miR-34b和miR-34c。它们在不同转移潜能的前列腺癌细胞中的表达具有明显差异，其中miR-34b和miR-34c差异显著。
发明内容
本发明的目的是：发现了miR-34b和miR-34c具有抑制前列腺癌转移作用，为前列腺癌转移治疗提供新的生物标志物。
本发明的技术方案是：MicroRNA-34家族成员miR-34b作为抑制转移性前列腺癌细胞迁移和侵袭，抑制癌症转移的应用。
MicroRNA-34家族成员miR-34c作为抑制转移性前列腺癌细胞迁移和侵袭，抑制癌症转移的应用，。
MicroRNA-34家族成员miR-34b和miR-34c联合使用作为抑制转移性前列腺癌细胞迁移和侵袭，抑制癌症转移的应用。
MicroRNA-34家族成员miR-34b和miR-34c其中之一或联合使用为治疗转移性前列腺癌提供生物学标志物。
本发明的过程是
1. 检测miR-34b和miR-34c在高转移潜能前列腺癌细胞PC-3和低转移潜能前列腺癌细胞DU145细胞中的表达量。
2. 转染mimic及inhibitor 高表达和抑制前列腺癌细胞中miR-34b和miR-34c。
3. 利用Transwell实验检测高表达和低表达miR-34b和miR-34c的前列腺癌细胞的迁移和侵袭能力。
本发明的有益效果是：miR-34b和miR-34c抑制转移性前列腺癌细胞迁移和侵袭。为治疗转移性前列腺癌提供新的生物学标志物。
附图说明
附图1为实时荧光定量检测miR-34b和miR-34c在前列腺癌细胞PC-3、DU145中的表达量。
附图2为转染miR-34b/c mimic 和NC及未转染的PC-3细胞的迁移及侵袭能力。
附图3为图2中迁移和侵袭的细胞数量的统计。
附图4为转染miR-34b/c inhibitor 和NC及未转染的DU145细胞的迁移及侵袭能力。
附图5 图4中迁移和侵袭的细胞数量的统计。
具体实施方式
本发明的实施例：
Transwell法检测miR-34b和miR-34c影响前列腺癌细胞迁移和侵袭能力
细胞及其它试剂
人前列腺癌细胞株PC-3、DU145来自加拿大多伦多Sunnybrook 研究中心；DMEM培养基及胰酶购于美国Hyclone公司；胎牛血清（FBS）购于浙江天杭生物科技有限公司；TRIzol®reagent 、Lipofectamine® 3000 reagent购于美国Invitrogen公司；引物、mimic及inhibitor购自中国广州锐博生物科技有限公司；cDNA逆转录试剂盒、UItraSYBR GreenqPCR Mixture 购于康为世纪公司；Opti-MEM培养基购于美国Gibco公司；transwell 购于美国康宁公司；Matrigel 购于美国BD Biosciences公司。
细胞迁移检测
1. miR-34b/c 表达检测
1.1 培养PC-3及DU145细胞，收集细胞用TRIzol法提取总RNA，使用cDNA逆转录试剂盒将RNA逆转录为cDNA，用于PCR检测。
1.2 实时荧光定量检测miR-34b/c在PC-3及DU145细胞中的表达量。
2. 转染miR-34b/c mimic、inhibitor及对照Negative Control （NC）至前列腺癌细胞
使用Lipofectamine® 3000 reagent将miR-34b/c mimic及NC转染至PC-3细胞；miR-34b/c inhibitor及NC转染至DU145细胞中。转染时将Lipofectamine® 3000与mimic（50nM）及inhibitor（100nM）混合入Opti-MEM培养基中形成转染复合物，加入至含Opti-MEM培养基的细胞中转染48h。收集细胞提取总RNA，逆转录成cDNA进行实时荧光定量PCR检测转染效率。
3. Transwell 法检测细胞迁移和侵袭能力
收集转染mimic/inhibitor及NC48h细胞及未转染的细胞三组，消化并悬浮细胞，每组取1×105/ml细胞稀释于200μl Opti-MEM培养基接种于Transwell板每孔上室中，下室中加入700μl含10%FBS DMEM培养基，置37℃培养箱中培养14h。取出Transwell小室用75%乙醇固定细胞，0.1%结晶紫染色后用倒置相差显微镜观察迁移至下室的细胞数确定其迁移能力。检测细胞侵袭能力需在接种细胞前将上室内铺入200μl 1:5稀释的Matrigel。
4. 结果显示：
miR-34b/c 在高转移性前列腺癌细胞PC-3中低表达，在低转移性前列腺癌细胞DU145细胞中高表达。转染miR-34b/c mimic 的PC-3细胞比转染NC对照及未转染的细胞迁移和侵袭力弱；转染miR-34b/c inhibitor 的DU145细胞比转染NC对照及未转染的细胞迁移和侵袭力强。因此miR-34b/c具有抑制前列腺癌细胞迁移和侵袭的作用。
Claims (4)
1.MicroRNA-34家族成员miR-34b作为抑制转移性前列腺癌细胞迁移和侵袭，抑制癌症转移的应用。
2.MicroRNA-34家族成员miR-34c作为抑制转移性前列腺癌细胞迁移和侵袭，抑制癌症转移的应用，。
3.MicroRNA-34家族成员miR-34b和miR-34c联合使用作为抑制转移性前列腺癌细胞迁移和侵袭，抑制癌症转移的应用。
4.MicroRNA-34家族成员miR-34b和miR-34c其中之一或联合使用为治疗转移性前列腺癌提供生物学标志物。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711370384.6A CN108018354A (zh) | 2017-12-19 | 2017-12-19 | MicroRNA-34在抑制前列腺癌转移治疗中的新用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711370384.6A CN108018354A (zh) | 2017-12-19 | 2017-12-19 | MicroRNA-34在抑制前列腺癌转移治疗中的新用途 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108018354A true CN108018354A (zh) | 2018-05-11 |
Family
ID=62074130
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711370384.6A Withdrawn CN108018354A (zh) | 2017-12-19 | 2017-12-19 | MicroRNA-34在抑制前列腺癌转移治疗中的新用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108018354A (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110484605A (zh) * | 2019-09-23 | 2019-11-22 | 中国人民解放军陆军军医大学第一附属医院 | 一种活细胞原位检测microRNA-34的基因及其制备方法和应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101801419A (zh) * | 2007-06-08 | 2010-08-11 | 米尔纳疗法公司 | 作为治疗干预的靶标的miR-34调控的基因和路径 |
| CN102027129A (zh) * | 2008-02-28 | 2011-04-20 | 俄亥俄州立大学研究基金会 | 用于前列腺相关病症的诊断、预后和治疗的基于微rna的方法和组合物 |
| CN105473742A (zh) * | 2013-06-24 | 2016-04-06 | 米尔纳疗法公司 | miR-34活性的生物标志物 |
| US20160362689A1 (en) * | 2012-08-29 | 2016-12-15 | City Of Hope | Differentially expressed microrna molecules for the treatment and diagnosis of cancer |
-
2017
- 2017-12-19 CN CN201711370384.6A patent/CN108018354A/zh not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101801419A (zh) * | 2007-06-08 | 2010-08-11 | 米尔纳疗法公司 | 作为治疗干预的靶标的miR-34调控的基因和路径 |
| CN102027129A (zh) * | 2008-02-28 | 2011-04-20 | 俄亥俄州立大学研究基金会 | 用于前列腺相关病症的诊断、预后和治疗的基于微rna的方法和组合物 |
| US20160362689A1 (en) * | 2012-08-29 | 2016-12-15 | City Of Hope | Differentially expressed microrna molecules for the treatment and diagnosis of cancer |
| CN105473742A (zh) * | 2013-06-24 | 2016-04-06 | 米尔纳疗法公司 | miR-34活性的生物标志物 |
Non-Patent Citations (3)
| Title |
|---|
| BEATRIZ A WALTER等: "Comprehensive microRNA Profiling of Prostate Cancer", 《JOURNAL OF CANCER》 * |
| SHAHANA MAJID等: "miRNA-34b Inhibits Prostate Cancer through Demethylation, Active Chromatin Modifications, and AKT Pathways", 《CLIN CANCER RES》 * |
| 刘浩等: "miRNA一34e对人前列腺癌PC3细胞增殖和侵袭能力的影响", 《中国医师杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110484605A (zh) * | 2019-09-23 | 2019-11-22 | 中国人民解放军陆军军医大学第一附属医院 | 一种活细胞原位检测microRNA-34的基因及其制备方法和应用 |
| CN110484605B (zh) * | 2019-09-23 | 2020-07-28 | 中国人民解放军陆军军医大学第一附属医院 | 一种活细胞原位检测microRNA-34的基因及其制备方法和应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Integrated analysis of ascites and plasma extracellular vesicles identifies a miRNA-based diagnostic signature in ovarian cancer | |
| Kurashige et al. | MicroRNA-200b regulates cell proliferation, invasion, and migration by directly targeting ZEB2 in gastric carcinoma | |
| Fan et al. | miR‐122 promotes metastasis of clear‐cell renal cell carcinoma by downregulating Dicer | |
| Wang et al. | Loss of miR-100 enhances migration, invasion, epithelialmesenchymal transition and stemness properties in prostate cancer cells through targeting Argonaute 2 | |
| Tatarano et al. | Novel oncogenic function of mesoderm development candidate 1 and its regulation by MiR-574-3p in bladder cancer cell lines | |
| Wang et al. | HOXA11‐AS regulates JAK‐STAT pathway by miR‐15a‐3p/STAT3 axis to promote the growth and metastasis in liver cancer | |
| CN107805663B (zh) | Lnc03729基因作为生物标志物在肺腺癌预诊断试剂中的应用 | |
| Ahmad et al. | miR‐22 regulates expression of oncogenic neuro‐epithelial transforming gene 1, NET 1 | |
| US10604770B2 (en) | Method for extracting differentiated cells | |
| Budkova et al. | Expression of ncRNAs on the DLK1-DIO3 locus is associated with basal and mesenchymal phenotype in breast epithelial progenitor cells | |
| Liu et al. | MicroRNA‑19b promotes the migration and invasion of ovarian cancer cells by inhibiting the PTEN/AKT signaling pathway Retraction in/10.3892/ol. 2024.14424 | |
| Yang et al. | MiR-491 attenuates cancer stem cells-like properties of hepatocellular carcinoma by inhibition of GIT-1/NF-κB-mediated EMT | |
| Huang et al. | miR-888 in MCF-7 side population sphere cells directly targets E-cadherin | |
| Lin et al. | MicroRNA-328 inhibits migration and epithelial–mesenchymal transition by targeting CD44 in nasopharyngeal carcinoma cells | |
| Li et al. | miR‑205 targets YAP1 and inhibits proliferation and invasion in thyroid cancer cells | |
| CN111718995A (zh) | 一种鼻咽癌转移诊断和/或预后评估的生物标记 | |
| Xiang et al. | Myocardin inhibits estrogen receptor alpha‐mediated proliferation of human breast cancer MCF‐7 cells via regulating MicroRNA expression | |
| CN104293788A (zh) | 抑制S100A9基因表达的siRNA及其应用 | |
| CN108018354A (zh) | MicroRNA-34在抑制前列腺癌转移治疗中的新用途 | |
| Ratnadiwakara et al. | SRSF3 shapes the structure of miR‐17‐92 cluster RNA and promotes selective processing of miR‐17 and miR‐20a | |
| Zheng et al. | Mir-382 promotes differentiation of rat liver progenitor cell WB-F344 by targeting Ezh2 | |
| Liu et al. | MicroRNA-204-3p inhibits metastasis of pancreatic cancer via downregulating MGAT1 | |
| Tang et al. | Circ_0006220 contributes to NSCLC progression through miR-342-3p/GOT2 Axis | |
| Ware et al. | Induction of mesenchymal-epithelial transitions in sarcoma cells | |
| Xu et al. | miR‑3120‑5p acts as a diagnostic biomarker in non‑small cell lung cancer and promotes cancer cell proliferation and invasion by targeting KLF4 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20180511 |