miR-34a mediates oxaliplatin resistance of colon cancer cells by inhibiting autophagy via transforming growth factor-β/Smad4 pathway_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

YAO Lei , LIN Yujia , GAO Shuang , JIANG Mingshan , MA Donglai , WANG Fujing ,  QIAO Pengfei

Department of General Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin 150086, P. R. China;

Corresponding author：

QIAO Pengfei, Email: lunwenqpf@126.com

Keywords：

colon cancer; miR-34a; oxaliplatin; autophagy; transforming growth factor-β/Smad4 pathway

DOI：

10.7507/1007-9424.201708098

Video：

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Objective To investigate whether miRNA (miR)-34a mediates oxaliplatin (OXA) resistance of colon cancer cells by inhibiting macroautophagy via the transforming growth factor (TGF)-β/Smad4 pathway. Methods miR-34a expression levels were detected in colon cancer tissues and colon cancer cell lines by quantitative real-time polymerase chain reaction (qRT-PCR). Computational search, functional luciferase assay, and Western blotting method were used to demonstrate the downstream target of miR-34a in colon cancer cells. Cell viability was measured with cell counting kit-8. Apoptosis and macroautophagy of colon cancer cells were analyzed by flow cytometry and transmission electron microscopy, and expressions of Beclin1 and LC3Ⅱ protein were detected by Western blotting method. Results Expression of miR-34a was significantly reduced while expressions of TGF-β and Smad4 mRNA were increased in colon cancer patients treated with OXA-based chemotherapy. OXA treatment also resulted in decreased miR-34a expression levels and increased TGF-β and Smad4 expression levels in both parental cells and the OXA-resistant colon cancer cells. Activation of macroautophagy contributed to OXA resistance in colon cancer cells. Expression levels of Smad4 and miR-34a in colon cancer patients had a significant inverse correlation and overexpressing miR-34a inhibited macroautophagy activation by directly targeting Smad4 through the TGF-β/Smad4 pathway. OXA-induced downregulation of miR-34a and increased drug resistance by activating macroautophagy in colon cancer cells. Conclusion miR-34a mediates OXA resistance of colon cancer by inhibiting autophagy via the TGF-β/Smad4 pathway.

Citation： YAO Lei, LIN Yujia, GAO Shuang, JIANG Mingshan, MA Donglai, WANG Fujing, QIAO Pengfei. miR-34a mediates oxaliplatin resistance of colon cancer cells by inhibiting autophagy via transforming growth factor-β/Smad4 pathway. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2018, 25(6): 680-688. doi: 10.7507/1007-9424.201708098 Copy

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14.	刘虹, 邵荣光. 自噬在肿瘤发生与发展过程中的调节作用. 药学学报, 2016, 51(1): 23-28.
15.	Edinger AL, Thompson CB. Death by design: apoptosis, necrosis and autophagy. Curr Opin Cell Biol, 2004, 16(6): 663-669.
16.	Zhao Z, Han F, Yang S, et al. Oxamate-mediated inhibition of lactate dehydrogenase induces protective autophagy in gastric cancer cells: involvement of the Akt-mTOR signaling pathway. Cancer Lett, 2015, 358(1): 17-26.
17.	Lv L, Liu HG, Dong SY, et al. Upregulation of CD44v6 contributes to acquired chemoresistance via the modulation of autophagy in colon cancer SW480 cells. Tumour Biol, 2016, 37(7): 8811-8824.
18.	Shi Y, Han Y, Xie F, et al. ASPP2 enhances oxaliplatin (L-OHP)-induced colorectal cancer cell apoptosis in a p53-independent manner by inhibiting cell autophagy. J Cell Mol Med, 2015, 19(3): 535-543.
19.	Xu T, Zhu Y, Xiong Y, et al. MicroRNA-195 suppresses tumorigenicity and regulates G1/S transition of human hepatocellular carcinoma cells. Hepatology, 2009, 50(1): 113-121.
20.	Nam EJ, Kim S, Lee TS, et al. Primary and recurrent ovarian high-grade serous carcinomas display similar microRNA expression patterns relative to those of normal ovarian tissue. Oncotarget, 2016, 7(43): 70524-70534.
21.	Li X, Zhao H, Zhou X, et al. Inhibition of lactate dehydrogenase A by microRNA-34a resensitizes colon cancer cells to 5-fluorouracil. Mol Med Rep, 2014, 11(1): 577-582.
22.	Li H, Rokavec M, Hermeking H. Soluble IL6R represents a miR-34a target: potential implications for the recently identified IL-6R/STAT3/miR-34a feed-back loop. Oncotarget, 2015, 6(16): 14026-14032.
23.	El-Tanbouly DM, Wadie W, Sayed RH. Modulation of TGF-β/Smad and ERK signaling pathways mediates the anti-fibrotic effect of mirtazapine in mice. Toxicol Appl Pharmacol, 2017, 329: 224-230.

1. Ung L, Lam AK, Morris DL, et al. Tissue-based biomarkers predicting outcomes in metastatic colorectal cancer: a review. Clin Transl Oncol, 2014, 16(5): 425-435.
2. Chen W, Zheng R, Baade PD, et al. Cancer statistics in China, 2015. CA Cancer J Clin, 2016, 66(2): 115-132.
3. Cuccurullo V. AJCC Cancer Staging Handbook: from the AJCC Cancer Staging Manual (7^th edition). European J Nuclear Med Molec Imag, 2011, 38(2): 408-408.
4. Compton CC. Optimal pathologic staging: defining stageⅡ disease. Clin Cancer Res, 2007, 13(22 Pt 2): 6862s-6870s.
5. 张升, 蔡国响, 蔡三军. 多学科综合治疗与精准医学治疗用于结直肠癌诊治的探讨. 上海医药, 2016, 37(23): 18-20.
6. Yoo PS, Lopez-Soler RI, Longo WE, et al. Liver resection for metastatic colorectal cancer in the age of neoadjuvant chemotherapy and bevacizumab. Clin Colorectal Cancer, 2006, 6(3): 202-207.
7. 付强, 张永磊, 陈小兵, 等. MiR-145 对人结肠癌细胞株 HCT-116 奥沙利铂耐药机制探讨. 中华肿瘤防治杂志, 2015, 22(19): 1523-1527.
8. Peraltazaragoza O, Deas J, Menesesacosta A, et al. Relevance of miR-21 in regulation of tumor suppressor gene PTEN in human cervical cancer cells. Bmc Cancer, 2016, 16(1): 215.
9. Zhou JY, Chen X, Zhao J, et al. MicroRNA-34a overcomes HGF-mediated gefitinib resistance in EGFR mutant lung cancer cells partly by targeting MET. Cancer Lett, 2014, 351(2): 265-271.
10. Qiao P, Li G, Bi W, et al. microRNA-34a inhibits epithelial mesenchymal transition in human cholangiocarcinoma by targeting Smad4 through transforming growth factor-beta/Smad pathway. BMC Cancer, 2015, 15: 469.
11. Yang HZ, Ma Y, Zhou Y, et al. Autophagy contributes to the enrichment and survival of colorectal cancer stem cells under oxaliplatin treatment. Cancer Lett, 2015, 361(1): 128-136.
12. Suzuki HI, Kiyono K, Miyazono K. Regulation of autophagy by transforming growth factor-β (TGF-β) signaling. Autophagy, 2010, 6(5): 645-647.
13. 祝丹希, 孟祥军. miRNA 与结肠癌化疗敏感性关系的研究进展. 胃肠病学和肝病学杂志, 2016, 25(9): 965-967.
14. 刘虹, 邵荣光. 自噬在肿瘤发生与发展过程中的调节作用. 药学学报, 2016, 51(1): 23-28.
15. Edinger AL, Thompson CB. Death by design: apoptosis, necrosis and autophagy. Curr Opin Cell Biol, 2004, 16(6): 663-669.
16. Zhao Z, Han F, Yang S, et al. Oxamate-mediated inhibition of lactate dehydrogenase induces protective autophagy in gastric cancer cells: involvement of the Akt-mTOR signaling pathway. Cancer Lett, 2015, 358(1): 17-26.
17. Lv L, Liu HG, Dong SY, et al. Upregulation of CD44v6 contributes to acquired chemoresistance via the modulation of autophagy in colon cancer SW480 cells. Tumour Biol, 2016, 37(7): 8811-8824.
18. Shi Y, Han Y, Xie F, et al. ASPP2 enhances oxaliplatin (L-OHP)-induced colorectal cancer cell apoptosis in a p53-independent manner by inhibiting cell autophagy. J Cell Mol Med, 2015, 19(3): 535-543.
19. Xu T, Zhu Y, Xiong Y, et al. MicroRNA-195 suppresses tumorigenicity and regulates G1/S transition of human hepatocellular carcinoma cells. Hepatology, 2009, 50(1): 113-121.
20. Nam EJ, Kim S, Lee TS, et al. Primary and recurrent ovarian high-grade serous carcinomas display similar microRNA expression patterns relative to those of normal ovarian tissue. Oncotarget, 2016, 7(43): 70524-70534.
21. Li X, Zhao H, Zhou X, et al. Inhibition of lactate dehydrogenase A by microRNA-34a resensitizes colon cancer cells to 5-fluorouracil. Mol Med Rep, 2014, 11(1): 577-582.
22. Li H, Rokavec M, Hermeking H. Soluble IL6R represents a miR-34a target: potential implications for the recently identified IL-6R/STAT3/miR-34a feed-back loop. Oncotarget, 2015, 6(16): 14026-14032.
23. El-Tanbouly DM, Wadie W, Sayed RH. Modulation of TGF-β/Smad and ERK signaling pathways mediates the anti-fibrotic effect of mirtazapine in mice. Toxicol Appl Pharmacol, 2017, 329: 224-230.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

miR-34a mediates oxaliplatin resistance of colon cancer cells by inhibiting autophagy via transforming growth factor-β/Smad4 pathway

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