Concomitant Inhibition of Cytoprotective Autophagy Augments the Efficacy of Withaferin A in Hepatocellular Carcinoma
<p>Withaferin A inhibits growth and clonogenicity of HCC cells. (<b>A</b>) Cell viability of Huh7, HepG2, MHCC97H and MHCC97L cells was examined using MTT assay after treatment with indicated concentrations of WFA compared to respective vehicle treated controls (denoted by “C”). (<b>B</b>) Huh7, HepG2, MHCC97H and MHCC97L cells were treated with mentioned concentrations of WFA followed by trypan blue dye exclusion assay. (<b>C</b>) Huh7 and MHCC97H cells were treated with indicated concentrations of WFA and subjected to clonogenicity assay. * <span class="html-italic">p</span> < 0.05, compared with control; ** <span class="html-italic">p</span> < 0.01, compared with control; *** <span class="html-italic">p</span> < 0.001, compared with control.</p> "> Figure 2
<p>Withaferin A induces LC3B conversion and augments autophagy in vitro and in vivo. (<b>A</b>,<b>B</b>) Expression level of LC3B in HCC cells after treatment with 5 µM of WFA by immunoblotting. ACTB served as the loading control. (<b>C</b>) HCC cells were transfected with a GFP-tagged LC3B-encoding plasmid, followed by treatment with 5 µM WFA or EBSS. EBSS was used as a positive control for autophagy-induction. Representative images are shown. LC3B puncta were counted and shown as bar diagram. ** <span class="html-italic">p</span> < 0.01, compared with control; *** <span class="html-italic">p</span> < 0.001, compared with control. Scale bar 20 μm. (<b>D</b>) HepG2 derived <span class="html-italic">xenograft</span> tumors from vehicle-treated and WFA-treated mice were subjected to immunohistochemical analysis for autophagy related proteins (LC3B, ATG5, ATG7, BECN1 and SQSTM1). IHC signals were quantified using Aperio ImageScope Software, Leica and shown as bar graphs. * <span class="html-italic">p</span> < 0.05, compared with control; ** <span class="html-italic">p</span> < 0.01, compared with control; *** <span class="html-italic">p</span> < 0.001, compared with control. Scale bar 100 μm (<b>E</b>) HCC cells were treated with 5 μM WFA and immunoblotted for the expression level of autophagy related proteins (ATG5, ATG7, BECN1 and SQSTM1). ACTB served as the loading control. (<b>F</b>) Total RNA was extracted from the WFA treated and vehicle treated HCC cells and the expression of ATG5, ATG7 and BECN1 was studied. ACTB was used as the loading control.</p> "> Figure 3
<p>Withaferin A increases the autophagic flux in HCC cells. (<b>A</b>) Upper panel represent the schematic diagram of tfLC3B. Huh7 and MHCC97H cells were transfected with tfLC3B and treated with 5 µM WFA or EBSS. Representative images are shown. Red and yellow puncta were counted and shown as bar diagram. * <span class="html-italic">p</span> < 0.05, compared with control; ** <span class="html-italic">p</span> < 0.01, compared with control, Scale bar 10 μm. (<b>B</b>,<b>C</b>) Upper panel represent the schematic diagram of tfLC3B. Huh7 cells were transfected with tfLC3B and treated with 5 μM WFA in combination with 25 µM CQ. Representative images are shown. Red and yellow puncta were counted and shown as a bar diagram. ** <span class="html-italic">p</span> < 0.01, compared with control; *** <span class="html-italic">p</span> < 0.001, compared with control, Scale bar 10 μm (<b>D</b>) Huh7 and MHCC97H cells were treated with 5 μM WFA in combination with 25 µM CQ and 200 nM bafilomycin and subjected to immunoblot analyses for the expression level of LC3B. ACTB served as the loading control. (<b>E</b>–<b>G</b>) Huh7 and MHCC97H cells were treated with 5 μM WFA in combination with 25 µM CQ and 200 nM bafilomycin and subjected to immunofluorescence analyses for LC3B. Representative images are shown. Green puncta were counted and shown as bar diagram. *** <span class="html-italic">p</span> < 0.001, compared with WFA alone, Scale bar 10 μm.</p> "> Figure 4
<p>Withaferin A elevates the fusion of autophagosomes with lysosomes. (<b>A</b>) Huh7 and MHCC97H cells were transfected with GFP-LC3B encoding plasmid, treated with 5 µM of WFA or EBSS, followed by Lysotracker red staining. Representative fluorescence images are shown. Number of yellow puncta shown as bar graphs. ** <span class="html-italic">p</span> < 0.01, compared with control. (<b>B</b>) Huh7 and MHCC97H cells were co-transfected with GFP-LC3B and RFP-Rab7 followed by 5 μM WFA or EBSS treatment. Representative fluorescence images are shown here. Bar graphs show number of yellow puncta. ** <span class="html-italic">p</span> < 0.01, compared with control; *** <span class="html-italic">p</span> < 0.001, compared with control. Scale bar 10 μm.</p> "> Figure 5
<p>Withaferin A induces proteolytic degradation and Cathepsin D in lysosomes. (<b>A</b>,<b>B</b>) Huh7 and MHCC97H were treated with DQ-BSA followed by treatment with 5 µM of WFA or EBSS; fixed and stained with Lysotracker-Red. Representative images are shown. Bar diagrams show quantification of green and yellow puncta. * <span class="html-italic">p</span> < 0.05, compared with control; ** <span class="html-italic">p</span> < 0.01, compared with control; *** <span class="html-italic">p</span> < 0.001, compared with control. Scale bar 10 μm (<b>C</b>) Western blot analysis showing Cathepsin D and LAMP1 expression levels in the total cellular lysate and in the lysosomal extract of vehicle and WFA treated HUH7 and HepG2. (<b>D</b>,<b>E</b>) Cathepsin D activity assay using lysates from vehicle and WFA treated Huh7 and HepG2 cells.</p> "> Figure 6
<p>Withaferin A-mediated autophagy in HCC cells is cytoprotective in nature. (<b>A</b>–<b>D</b>) Cell viability of various HCC cells after treatment with 5 µM WFA alone and in combination with 4 mM of 3MA (3-Methyl adenine), 25 µM CQ or 200 nM bafilomycin, respectively. (<b>E</b>–<b>H</b>) HCC cells were treated with 5 µM WFA alone and in combination with 4 mM 3MA, 25 µM CQ and 200 nM bafilomycin, respectively, and subjected to trypan blue dye exclusion assay. * <span class="html-italic">p</span> < 0.05, compared with Withaferin A; ** <span class="html-italic">p</span> < 0.01, compared with WFA; *** <span class="html-italic">p</span> < 0.001, compared with Withaferin A.</p> "> Figure 7
<p>Synergistic interaction between Withaferin A and autophagic inhibitors. Inhibitory effects of WFA combined with 3MA, Chloroquine (CQ) and bafilomycin, respectively, in various HCC cells. The CI value was calculated using the Chou-Talalay method. CI < 1, CI = 1 and CI > 1 indicates synergistic, additive and antagonistic effects, respectively.</p> "> Figure 8
<p>Combined treatment with Withaferin A and autophagic inhibitors induces apoptosis in HCC cells. (<b>A</b>) HCC cells were treated with 5 µM WFA alone and in combination of 4 mM 3MA, 25 µM CQ and 200 nM bafilomycin, respectively, total cellular lysates were prepared and subjected to western blot analysis to examine the expression level of Cleaved-PARP (C-PARP) and Total-PARP (T-PARP), respectively. ACTB served as the loading control. (<b>B</b>) Huh7 and MHCC97H were treated with 5 µM WFA alone or co-treated with 4 mM 3MA, 25 µM CQ and 200 nM bafilomycin, respectively, and subjected to TUNNEL assay following the manufacturer’s protocol. Images were captured microscopically at 100× (Huh7) and 50× (MHCC97H) magnification. Bar graphs show number of apoptotic cells. ** <span class="html-italic">p</span> < 0.01, compared with WFA; *** <span class="html-italic">p</span> < 0.001, compared with WFA, Scale bar, 100 μm.</p> ">
Abstract
:1. Introduction
2. Results
2.1. Withaferin a Treatment Inhibits Viability and Clonogenicity of Hepatocellular Carcinoma Cells
2.2. Increased Conversion of LC3B upon Withaferin Treatment Indicates Autophagic Induction
2.3. Withaferin a Augments the Formation of Autophagosomes in Hepatocellular Carcinoma Cells
2.4. Induction of Lysosomal Activity Upon Withaferin a Treatment Exhibits a Functional Autophagic Response
2.5. Withaferin-Induced Autophagy is Cytoprotective in Function
2.6. Simultaneous Inhibition of Cytoprotective Autophagy Along with Withaferin a Treatment Synergistically Inhibits Hepatocellular Carcinoma Cells
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Reagents
4.2. MTT Cell Viability Assay
4.3. Trypan Blue Dye Exclusion Assay
4.4. Clonogenic Cell Survival Assay
4.5. Immunofluorescence Microscopy
4.6. Immunohistochemical Staining
4.7. Immunoblotting
4.8. Semi-Quantitative PCR
4.9. Cathepsin-D Activity Assay
4.10. TUNEL Assay for Apoptosis
4.11. Statistical Analysis
5. Conclusions
Author Contributions
Acknowledgments
Conflicts of Interest
References
- Forner, A.; Reig, M.; Bruix, J. Hepatocellular carcinoma. Lancet 2018, 391, 1301–1314. [Google Scholar] [CrossRef]
- Ferlay, J.; Soerjomataram, I.; Dikshit, R.; Eser, S.; Mathers, C.; Rebelo, M.; Parkin, D.M.; Forman, D.; Bray, F. Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int. J. Cancer 2015, 136, 9. [Google Scholar] [CrossRef]
- Byam, J.; Renz, J.; Millis, J.M. Liver transplantation for hepatocellular carcinoma. Hepatobiliary Surg. Nutr. 2013, 2, 22–30. [Google Scholar] [PubMed]
- Bruix, J.; Sherman, M. Management of hepatocellular carcinoma: An update. Hepatology 2011, 53, 1020–1022. [Google Scholar] [CrossRef] [Green Version]
- Llovet, J.M.; Ricci, S.; Mazzaferro, V.; Hilgard, P.; Gane, E.; Blanc, J.F.; de Oliveira, A.C.; Santoro, A.; Raoul, J.L.; Forner, A.; et al. Sorafenib in advanced hepatocellular carcinoma. N. Engl. J. Med. 2008, 359, 378–390. [Google Scholar] [CrossRef]
- Cheng, A.L.; Kang, Y.K.; Chen, Z.; Tsao, C.J.; Qin, S.; Kim, J.S.; Luo, R.; Feng, J.; Ye, S.; Yang, T.S.; et al. Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma: A phase III randomised, double-blind, placebo-controlled trial. Lancet Oncol. 2009, 10, 25–34. [Google Scholar] [CrossRef]
- Han, K.; Kim, J.H.; Ko, G.Y.; Gwon, D.I.; Sung, K.B. Treatment of hepatocellular carcinoma with portal venous tumor thrombosis: A comprehensive review. World J. Gastroenterol. 2016, 22, 407–416. [Google Scholar] [CrossRef] [PubMed]
- Chin, Y.W.; Balunas, M.J.; Chai, H.B.; Kinghorn, A.D. Drug discovery from natural sources. AAPS J. 2006, 8, E239–E253. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cragg, G.M.; Newman, D.J.; Snader, K.M. Natural products in drug discovery and development. J. Nat. Prod. 1997, 60, 52–60. [Google Scholar] [CrossRef]
- Ganesan, A. The impact of natural products upon modern drug discovery. Curr. Opin. Chem. Biol. 2008, 12, 306–317. [Google Scholar] [CrossRef] [PubMed]
- Patridge, E.; Gareiss, P.; Kinch, M.S.; Hoyer, D. An analysis of FDA-approved drugs: Natural products and their derivatives. Drug Discov. Today 2016, 21, 204–207. [Google Scholar] [CrossRef]
- Mantle, D.; Lennard, T.W.; Pickering, A.T. Therapeutic applications of medicinal plants in the treatment of breast cancer: A review of their pharmacology, efficacy and tolerability. Advers. Drug React. Toxicol. Rev. 2000, 19, 223–240. [Google Scholar]
- Mehbub, M.F.; Lei, J.; Franco, C.; Zhang, W. Marine sponge derived natural products between 2001 and 2010: Trends and opportunities for discovery of bioactives. Mar. Drugs 2014, 12, 4539–4577. [Google Scholar] [CrossRef] [PubMed]
- Mirjalili, M.H.; Moyano, E.; Bonfill, M.; Cusido, R.M.; Palazon, J. Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules 2009, 14, 2373–2393. [Google Scholar] [CrossRef]
- Vanden Berghe, W.; Sabbe, L.; Kaileh, M.; Haegeman, G.; Heyninck, K. Molecular insight in the multifunctional activities of Withaferin A. Biochem. Pharmacol. 2012, 84, 1282–1291. [Google Scholar] [CrossRef]
- Mishra, L.C.; Singh, B.B.; Dagenais, S. Scientific basis for the therapeutic use of Withania somnifera (ashwagandha): A review. Altern. Med. Rev. 2000, 5, 334–346. [Google Scholar] [PubMed]
- Misra, L.; Mishra, P.; Pandey, A.; Sangwan, R.S.; Sangwan, N.S.; Tuli, R. Withanolides from Withania somnifera roots. Phytochemistry 2008, 69, 1000–1004. [Google Scholar] [CrossRef]
- Chaurasiya, N.D.; Uniyal, G.C.; Lal, P.; Misra, L.; Sangwan, N.S.; Tuli, R.; Sangwan, R.S. Analysis of withanolides in root and leaf of Withania somnifera by HPLC with photodiode array and evaporative light scattering detection. Phytochem. Anal. 2008, 19, 148–154. [Google Scholar] [CrossRef]
- Chirumamilla, C.S.; Perez-Novo, C.; Van Ostade, X.; Vanden Berghe, W. Molecular insights into cancer therapeutic effects of the dietary medicinal phytochemical withaferin A. Proc. Nutr. Soc. 2017, 76, 96–105. [Google Scholar] [CrossRef]
- Yan, Z.; Guo, R.; Gan, L.; Lau, W.B.; Cao, X.; Zhao, J.; Ma, X.; Christopher, T.A.; Lopez, B.L.; Wang, Y. Withaferin A inhibits apoptosis via activated Akt-mediated inhibition of oxidative stress. Life Sci. 2018, 211, 91–101. [Google Scholar] [CrossRef]
- Chandrasekaran, B.; Pal, D.; Kolluru, V.; Tyagi, A.; Baby, B.; Dahiya, N.R.; Youssef, K.; Alatassi, H.; Ankem, M.K.; Sharma, A.K.; et al. The chemopreventive effect of withaferin A on spontaneous and inflammation-associated colon carcinogenesis models. Carcinogenesis 2018, 39, 1537–1547. [Google Scholar] [CrossRef] [PubMed]
- Xia, S.; Miao, Y.; Liu, S. Withaferin A induces apoptosis by ROS-dependent mitochondrial dysfunction in human colorectal cancer cells. Biochem. Biophys. Res. Commun. 2018, 503, 2363–2369. [Google Scholar] [CrossRef] [PubMed]
- Samanta, S.K.; Lee, J.; Hahm, E.R.; Singh, S.V. Peptidyl-prolyl cis/trans isomerase Pin1 regulates withaferin A-mediated cell cycle arrest in human breast cancer cells. Mol. Carcinog. 2018, 57, 936–946. [Google Scholar] [CrossRef] [PubMed]
- Chang, H.W.; Li, R.N.; Wang, H.R.; Liu, J.R.; Tang, J.Y.; Huang, H.W.; Chan, Y.H.; Yen, C.Y. Withaferin A Induces Oxidative Stress-Mediated Apoptosis and DNA Damage in Oral Cancer Cells. Front. Physiol. 2017, 8, 634. [Google Scholar] [CrossRef] [PubMed]
- Aliebrahimi, S.; Kouhsari, S.M.; Arab, S.S.; Shadboorestan, A.; Ostad, S.N. Phytochemicals, withaferin A and carnosol, overcome pancreatic cancer stem cells as c-Met inhibitors. Biomed. Pharm. 2018, 106, 1527–1536. [Google Scholar] [CrossRef]
- Kyakulaga, A.H.; Aqil, F.; Munagala, R.; Gupta, R.C. Withaferin A inhibits Epithelial to Mesenchymal Transition in Non-Small Cell Lung Cancer Cells. Sci. Rep. 2018, 8, 15737. [Google Scholar] [CrossRef] [PubMed]
- Nagalingam, A.; Kuppusamy, P.; Singh, S.V.; Sharma, D.; Saxena, N.K. Mechanistic elucidation of the antitumor properties of withaferin a in breast cancer. Cancer Res. 2014, 74, 2617–2629. [Google Scholar] [CrossRef]
- Kuppusamy, P.; Nagalingam, A.; Muniraj, N.; Saxena, N.K.; Sharma, D. Concomitant activation of ETS-like transcription factor-1 and Death Receptor-5 via extracellular signal-regulated kinase in withaferin A-mediated inhibition of hepatocarcinogenesis in mice. Sci. Rep. 2017, 7, 17943. [Google Scholar] [CrossRef] [Green Version]
- Chengappa, K.N.R.; Brar, J.S.; Gannon, J.M.; Schlicht, P.J. Adjunctive Use of a Standardized Extract of Withania somnifera (Ashwagandha) to Treat Symptom Exacerbation in Schizophrenia: A Randomized, Double-Blind, Placebo-Controlled Study. J. Clin. Psychiatry 2018, 79. [Google Scholar] [CrossRef]
- Huang, F.; Wang, B.R.; Wang, Y.G. Role of autophagy in tumorigenesis, metastasis, targeted therapy and drug resistance of hepatocellular carcinoma. World J. Gastroenterol. 2018, 24, 4643–4651. [Google Scholar] [CrossRef]
- Akkoc, Y.; Gozuacik, D. Autophagy and liver cancer. Turk. J. Gastroenterol. 2018, 29, 270–282. [Google Scholar] [CrossRef] [PubMed]
- Green, D.R.; Levine, B. To be or not to be? How selective autophagy and cell death govern cell fate. Cell 2014, 157, 65–75. [Google Scholar] [CrossRef] [PubMed]
- Galluzzi, L.; Baehrecke, E.H.; Ballabio, A.; Boya, P.; Bravo-San Pedro, J.M.; Cecconi, F.; Choi, A.M.; Chu, C.T.; Codogno, P.; Colombo, M.I.; et al. Molecular definitions of autophagy and related processes. EMBO J. 2017, 36, 1811–1836. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kriel, J.; Loos, B. The good, the bad and the autophagosome: Exploring unanswered questions of autophagy-dependent cell death. Cell Death Differ. 2019. [Google Scholar] [CrossRef] [PubMed]
- Mizushima, N.; Yoshimori, T.; Levine, B. Methods in mammalian autophagy research. Cell 2010, 140, 313–326. [Google Scholar] [CrossRef] [PubMed]
- Kimura, S.; Noda, T.; Yoshimori, T. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 2007, 3, 452–460. [Google Scholar] [CrossRef] [PubMed]
- Guerra, F.; Bucci, C. Multiple Roles of the Small GTPase Rab7. Cells 2016, 5, 34. [Google Scholar] [CrossRef] [PubMed]
- Hasui, K.; Wang, J.; Jia, X.; Tanaka, M.; Nagai, T.; Matsuyama, T.; Eizuru, Y. Enhanced Autophagy and Reduced Expression of Cathepsin D Are Related to Autophagic Cell Death in Epstein-Barr Virus-Associated Nasal Natural Killer/T-Cell Lymphomas: An Immunohistochemical Analysis of Beclin-1, LC3, Mitochondria (AE-1), and Cathepsin D in Nasopharyngeal Lymphomas. Acta Histochem. Cytochem. 2011, 44, 119–131. [Google Scholar] [Green Version]
- Gewirtz, D.A. When cytoprotective autophagy isn’t… and even when it is. Autophagy 2014, 10, 391–392. [Google Scholar] [CrossRef] [Green Version]
- Gewirtz, D.A. The four faces of autophagy: Implications for cancer therapy. Cancer Res. 2014, 74, 647–651. [Google Scholar] [CrossRef]
- Tsukada, M.; Ohsumi, Y. Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae. FEBS Lett. 1993, 333, 169–174. [Google Scholar] [CrossRef] [Green Version]
- Li, L.; Liu, X.; Zhou, L.; Wang, W.; Liu, Z.; Cheng, Y.; Li, J.; Wei, H. Autophagy Plays a Critical Role in Insulin Resistance- Mediated Chemoresistance in Hepatocellular Carcinoma Cells by Regulating the ER Stress. J. Cancer 2018, 9, 4314–4324. [Google Scholar] [CrossRef]
- Sharma, K.; Le, N.; Alotaibi, M.; Gewirtz, D.A. Cytotoxic autophagy in cancer therapy. Int. J. Mol. Sci. 2014, 15, 10034–10051. [Google Scholar] [CrossRef]
- Chung, S.J.; Nagaraju, G.P.; Nagalingam, A.; Muniraj, N.; Kuppusamy, P.; Walker, A.; Woo, J.; Gyorffy, B.; Gabrielson, E.; Saxena, N.K.; et al. ADIPOQ/adiponectin induces cytotoxic autophagy in breast cancer cells through STK11/LKB1-mediated activation of the AMPK-ULK1 axis. Autophagy 2017, 13, 1386–1403. [Google Scholar] [CrossRef] [PubMed]
- Sharma, K.; Goehe, R.W.; Di, X.; Hicks, M.A., 2nd; Torti, S.V.; Torti, F.M.; Harada, H.; Gewirtz, D.A. A novel cytostatic form of autophagy in sensitization of non-small cell lung cancer cells to radiation by vitamin D and the vitamin D analog, EB 1089. Autophagy 2014, 10, 2346–2361. [Google Scholar] [CrossRef] [PubMed]
- Muniraj, N.; Siddharth, S.; Nagalingam, A.; Walker, A.; Woo, J.; Gyorffy, B.; Gabrielson, E.; Saxena, N.K.; Sharma, D. Withaferin A inhibits lysosomal activity to block autophagic flux and induces apoptosis via energetic impairment in breast cancer cells. Carcinogenesis 2019. [Google Scholar] [CrossRef] [PubMed]
- Yan, Z.; Su, G.; Gao, W.; He, J.; Shen, Y.; Zeng, Y.; Liu, X. Fluid shear stress induces cell migration and invasion via activating autophagy in HepG2 cells. Cell Adhes. Migr. 2019. [Google Scholar] [CrossRef] [PubMed]
- Zhang, X.; Jin, L.; Tian, Z.; Wang, J.; Yang, Y.; Liu, J.; Chen, Y.; Hu, C.; Chen, T.; Zhao, Y.; et al. Nitric oxide inhibits autophagy and promotes apoptosis in hepatocellular carcinoma. Cancer Sci. 2019. [Google Scholar] [CrossRef]
- Helmy, S.A.; El-Mesery, M.; El-Karef, A.; Eissa, L.A.; El Gayar, A.M. Chloroquine upregulates TRAIL/TRAILR2 expression and potentiates doxorubicin anti-tumor activity in thioacetamide-induced hepatocellular carcinoma model. Chem. Biol. Interact. 2018, 279, 84–94. [Google Scholar] [CrossRef] [PubMed]
- Park, H.H.; Choi, S.W.; Lee, G.J.; Kim, Y.D.; Noh, H.J.; Oh, S.J.; Yoo, I.; Ha, Y.J.; Koo, G.B.; Hong, S.S.; et al. A formulated red ginseng extract inhibits autophagic flux and sensitizes to doxorubicin-induced cell death. J. Ginseng. Res. 2019, 43, 86–94. [Google Scholar] [CrossRef]
- Rong, L.W.; Wang, R.X.; Zheng, X.L.; Feng, X.Q.; Zhang, L.; Zhang, L.; Lin, Y.; Li, Z.P.; Wang, X. Combination of wogonin and sorafenib effectively kills human hepatocellular carcinoma cells through apoptosis potentiation and autophagy inhibition. Oncol. Lett. 2017, 13, 5028–5034. [Google Scholar] [CrossRef] [Green Version]
- Liu, W.; Yu, G.; Yu, W.; Ye, X.; Jin, Y.; Shrestha, A.; Yang, Q.; Sun, H. Autophagy Inhibits Apoptosis Induced by agrocybe aegerita Lectin in Hepatocellular Carcinoma. Anticancer Agents Med. Chem. 2017, 17, 221–229. [Google Scholar] [CrossRef] [PubMed]
- Zhong, J.; Dong, X.; Xiu, P.; Wang, F.; Liu, J.; Wei, H.; Xu, Z.; Liu, F.; Li, T.; Li, J. Blocking autophagy enhances meloxicam lethality to hepatocellular carcinoma by promotion of endoplasmic reticulum stress. Cell Prolif. 2015, 48, 691–704. [Google Scholar] [CrossRef] [PubMed]
- Cui, Z.; Zhang, Y.; Xia, K.; Yan, Q.; Kong, H.; Zhang, J.; Zuo, X.; Shi, J.; Wang, L.; Zhu, Y.; et al. Nanodiamond autophagy inhibitor allosterically improves the arsenical-based therapy of solid tumors. Nat. Commun. 2018, 9, 4347. [Google Scholar] [CrossRef] [PubMed]
- Haas, N.B.; Appleman, L.J.; Stein, M.; Redlinger, M.; Wilks, M.; Xu, X.; Onorati, A.; Kalavacharla, A.; Kim, T.; Zhen, C.J.; et al. Autophagy inhibition to augment mTOR inhibition: A phase I/II trial of everolimus and hydroxychloroquine in patients with previously treated renal cell carcinoma. Clin. Cancer Res. 2019. [Google Scholar] [CrossRef] [PubMed]
- Patel, S.; Hurez, V.; Nawrocki, S.T.; Goros, M.; Michalek, J.; Sarantopoulos, J.; Curiel, T.; Mahalingam, D. Vorinostat and hydroxychloroquine improve immunity and inhibit autophagy in metastatic colorectal cancer. Oncotarget 2016, 7, 59087–59097. [Google Scholar] [CrossRef]
- Avtanski, D.B.; Nagalingam, A.; Tomaszewski, J.E.; Risbood, P.; Difillippantonio, M.J.; Saxena, N.K.; Malhotra, S.V.; Sharma, D. Indolo-pyrido-isoquinolin based alkaloid inhibits growth, invasion and migration of breast cancer cells via activation of p53-miR34a axis. Mol. Oncol. 2016, 10, 1118–1132. [Google Scholar] [CrossRef]
- Avtanski, D.B.; Nagalingam, A.; Bonner, M.Y.; Arbiser, J.L.; Saxena, N.K.; Sharma, D. Honokiol activates LKB1-miR-34a axis and antagonizes the oncogenic actions of leptin in breast cancer. Oncotarget 2015, 6, 29947–29962. [Google Scholar] [CrossRef] [Green Version]
- Siddharth, S.; Goutam, K.; Das, S.; Nayak, A.; Nayak, D.; Sethy, C.; Wyatt, M.D.; Kundu, C.N. Nectin-4 is a breast cancer stem cell marker that induces WNT/beta-catenin signaling via Pi3k/Akt axis. Int. J. Biochem. Cell Biol. 2017, 89, 85–94. [Google Scholar] [CrossRef]
- Avtanski, D.B.; Nagalingam, A.; Bonner, M.Y.; Arbiser, J.L.; Saxena, N.K.; Sharma, D. Honokiol inhibits epithelial-mesenchymal transition in breast cancer cells by targeting signal transducer and activator of transcription 3/Zeb1/E-cadherin axis. Mol. Oncol. 2014, 8, 565–580. [Google Scholar] [CrossRef]
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Siddharth, S.; Muniraj, N.; Saxena, N.K.; Sharma, D. Concomitant Inhibition of Cytoprotective Autophagy Augments the Efficacy of Withaferin A in Hepatocellular Carcinoma. Cancers 2019, 11, 453. https://doi.org/10.3390/cancers11040453
Siddharth S, Muniraj N, Saxena NK, Sharma D. Concomitant Inhibition of Cytoprotective Autophagy Augments the Efficacy of Withaferin A in Hepatocellular Carcinoma. Cancers. 2019; 11(4):453. https://doi.org/10.3390/cancers11040453
Chicago/Turabian StyleSiddharth, Sumit, Nethaji Muniraj, Neeraj K. Saxena, and Dipali Sharma. 2019. "Concomitant Inhibition of Cytoprotective Autophagy Augments the Efficacy of Withaferin A in Hepatocellular Carcinoma" Cancers 11, no. 4: 453. https://doi.org/10.3390/cancers11040453