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Effect of lncRNA KCNQ1OT1 on autophagy and drug resistance of hepatocellular carcinoma cells by targeting miR-338-3p
Corresponding Author(s) : Qianqian Dai
Cellular and Molecular Biology,
Vol. 66 No. 3: Issue 3
Abstract
The current experiment aimed to investigate the effects of lncRNA KCNQ1OT1 on the proliferation, autophagy and drug resistance of hepatocellular carcinoma cells, as well as the potential molecular mechanism. Hepatocellular carcinoma SK-HEP-1 cells and DDP resistant SK-HEP-1/DDP cells were treated with cisplatin (DDP) of different concentrations (1 nmol/L, 2 nmol/L, 4 nmol/L, 8 nmol/L, 16 nmol/L). The survival rate of SK-HEP-1 and SK-HEP-1/DDP cells was determined by the CCK8 method. QRT-PCR was used to detect the levels of lncRNA KCNQ1OT1 and miR-338-3p in normal hepatocyte HH01, hepatocellular cell SK-HEP-1 and hepatoma cisplatin-resistant cell SK-HEP-1/DDP. Western blot was carried out to detect the expression levels of autophagy-related protein Beclin1 and proliferation-related protein P21 in cells. A dual-luciferase reporter assay system was performed to validate the relationship between KCNQ1OT1 and miR-338-3p. After the treatment of 1 nmol/L, 2 nmol/L, 4nmol/L, 8nmol/L and 16nmol/L cisplatin (DDP), the survival rate of SK-HEP-1/DDP cells is higher than that of SK-HEP-1 cells. The level of lncRNA KCNQ1OT1 was increased successively in HH01, SK-HEP-1 and SK-HEP-1/DDP cells, while miR-338-3p was decreased successively. Silencing lncRNA KCNQ1OT1 or over-expressing miR-338-3p combined with 16nmol/L DDP treatment reduced the survival rate of SK-HEP-1/DDP cells and up-regulate levels of P21 and Beclin1 proteins. LncRNA KCNQ1OT1 targeted and negatively regulated the expression of miR-338-3p. Inhibition of miR-338-3p reversed the effect of silencing lncRNA KCNQ1OT1 on survival, autophagy and cisplatin sensitivity of SK-HEP-1/DDP cell. LncRNA KCNQ1OT1 targets miR-338-3p to regulate the survival rate and autophagy of SK-HEP-1/DDP cells and improve the cisplatin sensitivity of SK-HEP-1/DDP cells. LncRNA KCNQ1OT1 is a potential molecular target for hepatocellular carcinoma.
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- Galluzzi L, Pietrocola F, Bravo"San Pedro JM, Amaravadi RK, Baehrecke EH, Cecconi F, Codogno P, Debnath J, Gewirtz DA, Karantza V, Kimmelman A. Autophagy in malignant transformation and cancer progression. EMBO J 2015; 34(7): 856-80.
- Huang F, Wang BR, Wang YG. Role of autophagy in tumorigenesis, metastasis, targeted therapy and drug resistance of hepatocellular carcinoma. World J Gastroenterol, 2018, 24(41):4643-4651.
- Fu Jun, Shang Haixu, Jia Hongzhi, et al. Relationship between Beclin1 and autophagy, tumors. Progress in Physiological Sciences, 2012, 43 (02): 77-80.
- Dou Chunqing, Jin Xin, Sun Liyuan, et al. miR-338-3p expression in hepatocellular carcinoma tissues and its relationship with clinicopathological parameters. Progress in Modern Biomedicine, 2017; 17 (12): 2290-2293.
- Zhao Jing, Wang Yuqing, Duan Siliang. Establishment of cisplatin-resistant human hepatocellular carcinoma cell line HepG2. J Qiqihar Med College, 2015; 36 (04): 477-478.
- Shan Yuying, Lu Caide. Research progress on the dual role of autophagy in hepatocellular carcinoma and the use of autophagy regulators in the treatment of hepatocellular carcinoma. Zhejiang Medical Journal, 2018; 40 (9): 1020-1023.
- Guan Wencai, Zhang Lingyun, Lin Qunbo, et al. Research progress on long-chain non-coding RNA in chemotherapy resistance of hepatocellular carcinoma. Chin Hepatol 2018; 23 (6): 543-545.
- Zheng L, Zhang FX, Wang LL, Hu HL, Lian YD. LncRNA KCNQ1OT1 is overexpressed in non-small cell lung cancer and its expression level is related to clinicopathology. Eur Rev Med Pharmacol Sci. 2019; 23(16):6944-50.
- Luo ZP, Jin H. Effects of LncRNA KCNQ1OT1 on proliferation and migration of ovarian cancer cells by Wnt/β-catenin. Eur Rev Med Pharmacol Sci, 2019, 23(20):8788-8794.
- Zhang S, Ma H, Zhang D, Xie S, Wang W, Li Q, Lin Z, Wang Y. LncRNA KCNQ1OT1 regulates proliferation and cisplatin resistance in tongue cancer via miR-211-5p mediated Ezrin/Fak/Src signaling. Cell death Dis 2018;9(7):1-6.
- Li Y, Li C, Li D, Yang L, Jin J, Zhang B. lncRNA KCNQ1OT1 enhances the chemoresistance of oxaliplatin in colon cancer by targeting the miR-34a/ATG4B pathway. OncoTargets Therapy. 2019; 12:2649.
- Xian D, Zhao Y. LncRNA KCNQ1OT1 enhanced the methotrexate resistance of colorectal cancer cells by regulating miR-760/PPP1R1B via the cAMP signalling pathway. J Cell Mol Med, 2019; 23(6):3808-3823.
- Hu H, Yang L, Li L, Zeng C. Long non-coding RNA KCNQ1OT1 modulates oxaliplatin resistance in hepatocellular carcinoma through miR-7-5p/ABCC1 axis. Biochemical and biophysical research communications. 2018; 503(4):2400-6.
- Wang Y, Qin H. miR-338-3p targets RAB23 and suppresses tumorigenicity of prostate cancer cells. Am J Cancer Res 2018; 8(12):2564-2574.
- Sun F, Yu M, Yu J, Liu Z, Zhou X, Liu Y, Ge X, Gao H, Li M, Jiang X, Liu S. miR-338-3p functions as a tumor suppressor in gastric cancer by targeting PTP1B. Cell Death Dis 2018;9(5):1-4.
- Vivacqua A, Sebastiani A, Miglietta AM, Rigiracciolo DC, Cirillo F, Galli GR, Talia M, Santolla MF, Lappano R, Giordano F, Panno ML. miR-338-3p is regulated by estrogens through GPER in breast cancer cells and Cancer-Associated Fibroblasts (CAFs). Cells. 2018;7(11): 203.
- Niu Q, Liu Z, Gao J, Wang Q. MiR-338-3p Enhances Ovarian Cancer Cell Sensitivity to Cisplatin by Downregulating WNT2B. Yonsei Med J. 2019; 60(12):1146-56.
- Han J, Li J, Tang K, Zhang H, Guo B, Hou N, Huang C. miR-338-3p confers 5-fluorouracil resistance in p53 mutant colon cancer cells by targeting the mammalian target of rapamycin. Exp Cell Res 2017;360(2):328-36.
- Xu H, Zhao L, Fang Q, Sun J, Zhang S, Zhan C, Liu S, Zhang Y. MiR-338-3p inhibits hepatocarcinoma cells and sensitizes these cells to sorafenib by targeting hypoxia-induced factor 1α. PloS one. 2014;9(12).
- Bordbar M, Darvishzadeh R, Pazhouhandeh M, Kahrizi D. An overview of genome editing methods based on endonucleases. Modern Genetics J 2020; 15(2): 75-92.
References
Galluzzi L, Pietrocola F, Bravo"San Pedro JM, Amaravadi RK, Baehrecke EH, Cecconi F, Codogno P, Debnath J, Gewirtz DA, Karantza V, Kimmelman A. Autophagy in malignant transformation and cancer progression. EMBO J 2015; 34(7): 856-80.
Huang F, Wang BR, Wang YG. Role of autophagy in tumorigenesis, metastasis, targeted therapy and drug resistance of hepatocellular carcinoma. World J Gastroenterol, 2018, 24(41):4643-4651.
Fu Jun, Shang Haixu, Jia Hongzhi, et al. Relationship between Beclin1 and autophagy, tumors. Progress in Physiological Sciences, 2012, 43 (02): 77-80.
Dou Chunqing, Jin Xin, Sun Liyuan, et al. miR-338-3p expression in hepatocellular carcinoma tissues and its relationship with clinicopathological parameters. Progress in Modern Biomedicine, 2017; 17 (12): 2290-2293.
Zhao Jing, Wang Yuqing, Duan Siliang. Establishment of cisplatin-resistant human hepatocellular carcinoma cell line HepG2. J Qiqihar Med College, 2015; 36 (04): 477-478.
Shan Yuying, Lu Caide. Research progress on the dual role of autophagy in hepatocellular carcinoma and the use of autophagy regulators in the treatment of hepatocellular carcinoma. Zhejiang Medical Journal, 2018; 40 (9): 1020-1023.
Guan Wencai, Zhang Lingyun, Lin Qunbo, et al. Research progress on long-chain non-coding RNA in chemotherapy resistance of hepatocellular carcinoma. Chin Hepatol 2018; 23 (6): 543-545.
Zheng L, Zhang FX, Wang LL, Hu HL, Lian YD. LncRNA KCNQ1OT1 is overexpressed in non-small cell lung cancer and its expression level is related to clinicopathology. Eur Rev Med Pharmacol Sci. 2019; 23(16):6944-50.
Luo ZP, Jin H. Effects of LncRNA KCNQ1OT1 on proliferation and migration of ovarian cancer cells by Wnt/β-catenin. Eur Rev Med Pharmacol Sci, 2019, 23(20):8788-8794.
Zhang S, Ma H, Zhang D, Xie S, Wang W, Li Q, Lin Z, Wang Y. LncRNA KCNQ1OT1 regulates proliferation and cisplatin resistance in tongue cancer via miR-211-5p mediated Ezrin/Fak/Src signaling. Cell death Dis 2018;9(7):1-6.
Li Y, Li C, Li D, Yang L, Jin J, Zhang B. lncRNA KCNQ1OT1 enhances the chemoresistance of oxaliplatin in colon cancer by targeting the miR-34a/ATG4B pathway. OncoTargets Therapy. 2019; 12:2649.
Xian D, Zhao Y. LncRNA KCNQ1OT1 enhanced the methotrexate resistance of colorectal cancer cells by regulating miR-760/PPP1R1B via the cAMP signalling pathway. J Cell Mol Med, 2019; 23(6):3808-3823.
Hu H, Yang L, Li L, Zeng C. Long non-coding RNA KCNQ1OT1 modulates oxaliplatin resistance in hepatocellular carcinoma through miR-7-5p/ABCC1 axis. Biochemical and biophysical research communications. 2018; 503(4):2400-6.
Wang Y, Qin H. miR-338-3p targets RAB23 and suppresses tumorigenicity of prostate cancer cells. Am J Cancer Res 2018; 8(12):2564-2574.
Sun F, Yu M, Yu J, Liu Z, Zhou X, Liu Y, Ge X, Gao H, Li M, Jiang X, Liu S. miR-338-3p functions as a tumor suppressor in gastric cancer by targeting PTP1B. Cell Death Dis 2018;9(5):1-4.
Vivacqua A, Sebastiani A, Miglietta AM, Rigiracciolo DC, Cirillo F, Galli GR, Talia M, Santolla MF, Lappano R, Giordano F, Panno ML. miR-338-3p is regulated by estrogens through GPER in breast cancer cells and Cancer-Associated Fibroblasts (CAFs). Cells. 2018;7(11): 203.
Niu Q, Liu Z, Gao J, Wang Q. MiR-338-3p Enhances Ovarian Cancer Cell Sensitivity to Cisplatin by Downregulating WNT2B. Yonsei Med J. 2019; 60(12):1146-56.
Han J, Li J, Tang K, Zhang H, Guo B, Hou N, Huang C. miR-338-3p confers 5-fluorouracil resistance in p53 mutant colon cancer cells by targeting the mammalian target of rapamycin. Exp Cell Res 2017;360(2):328-36.
Xu H, Zhao L, Fang Q, Sun J, Zhang S, Zhan C, Liu S, Zhang Y. MiR-338-3p inhibits hepatocarcinoma cells and sensitizes these cells to sorafenib by targeting hypoxia-induced factor 1α. PloS one. 2014;9(12).
Bordbar M, Darvishzadeh R, Pazhouhandeh M, Kahrizi D. An overview of genome editing methods based on endonucleases. Modern Genetics J 2020; 15(2): 75-92.