异丙酚对卵巢癌SKOV3细胞克隆形成、侵袭、EMT和miR-143表达的影响
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摘要
目的:探讨异丙酚对卵巢癌SKOV3细胞克隆形成、侵袭、EMT和miR-143表达的影响。方法:以0. 0、2. 5、5. 0和10. 0μmol/L异丙酚刺激SKOV3细胞48 h后,采用软琼脂集落形成实验检测细胞的克隆形成能力,Transwell小室实验检测细胞的侵袭能力,Western blot检测EMT相关蛋白E-cadherin和Vimentin的表达,实时荧光定量PCR检测细胞中miR-143的表达。脂质体法转染miR-143抑制剂后,观察下调miR-143表达对异丙酚刺激的SKOV3细胞的影响。结果:异丙酚能够呈浓度依赖性地抑制SKOV3细胞的克隆形成能力、侵袭能力和Vimentin蛋白的表达,促进E-cadherin蛋白和miR-143的表达(P <0. 05)。转染miR-143抑制剂后,异丙酚对SKOV3细胞的上述作用明显逆转(P <0. 05)。结论:异丙酚可能通过上调miR-143表达抑制SKOV3细胞克隆形成、侵袭和EMT进程,发挥抑制癌细胞转移的作用。
Aim: To investigate the effects of propofol on the cloning formation,invasive ability,epithelial mesenchymal transition,the expression of miR-143 of ovarian cancer SKOV3 cells and its mechanism. Methods: SKOV3 cells were stimulated with 0. 0,2. 5,5. 0 and 10. 0 μmol/L propofol for 48 hours. The colony forming ability of SKOV3 cells was detected by soft agar colony formation assay,the invasive ability of SKOV3 cells was detected by Transwell chamber assay,the expressions of EMT-related proteins E-cadherin and Vimentin were detected by Western blot,and the expression of miR-143 in SKOV3 cells was detected by real-time fluorescence quantitative PCR. The effects of down-regulation of the expression of miR-143 on propofol-stimulated SKOV3 cells were observed after transfection of miR-143 inhibitors by liposome method.Results: The cloning forming ability,invasive ability and Vimentin protein expression of SKOV3 cells were inhibited and the expressions of E-cadherin protein and miR-143 were promoted by propofol in a concentration-dependent manner( P <0. 05). The above effects of propofol on SKOV3 cells were reversed after transfection with the inhibitor of miR-143( P <0. 05). Conclusion: Propofol can inhibit SKOV3 cell clone formation,invasion and EMT process by up-regulating the expression of miR-143.
Aim: To investigate the effects of propofol on the cloning formation,invasive ability,epithelial mesenchymal transition,the expression of miR-143 of ovarian cancer SKOV3 cells and its mechanism. Methods: SKOV3 cells were stimulated with 0. 0,2. 5,5. 0 and 10. 0 μmol/L propofol for 48 hours. The colony forming ability of SKOV3 cells was detected by soft agar colony formation assay,the invasive ability of SKOV3 cells was detected by Transwell chamber assay,the expressions of EMT-related proteins E-cadherin and Vimentin were detected by Western blot,and the expression of miR-143 in SKOV3 cells was detected by real-time fluorescence quantitative PCR. The effects of down-regulation of the expression of miR-143 on propofol-stimulated SKOV3 cells were observed after transfection of miR-143 inhibitors by liposome method.Results: The cloning forming ability,invasive ability and Vimentin protein expression of SKOV3 cells were inhibited and the expressions of E-cadherin protein and miR-143 were promoted by propofol in a concentration-dependent manner( P <0. 05). The above effects of propofol on SKOV3 cells were reversed after transfection with the inhibitor of miR-143( P <0. 05). Conclusion: Propofol can inhibit SKOV3 cell clone formation,invasion and EMT process by up-regulating the expression of miR-143.
引文
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[14]LEI C,DU F,SUN LA,et al. miR-143 and miR-145 inhibit gastric cancer cell migration and metastasis by suppressing MYO6[J]. Cell Death Dis,2017,8(10):e3101
[15]WANG L,HE J,XU H,et al. MiR-143 targets CTGF and exerts tumor-suppressing functions in epithelial ovarian cancer[J]. Am J Transl Res,2016,8(6):2716
[16]HUANG X,TENG Y,YANG H,et al. Propofol inhibits invasion and growth of ovarian cancer cells via regulating miR-9/NF-kappa B signal[J]. Braz J Med and Biol Res,2016,49(12):e5717
[2] LIU SQ,ZHANG JL,LI ZW,et al. Propofol inhibits proliferation, migration, invasion and promotes apoptosis through down-regulating mir-374a in hepatocarcinoma cell lines[J]. Cell Physiol Biochem,2018,49(6):2099
[3] LIU ZM,ZHANG J,HONG GC,et al. Propofol inhibits growth and invasion of pancreatic cancer cells through regulation of the miR-21/Slug signaling pathway[J]. Am J Transl Res,2016,8(10):4120
[4] LI DF,HU JS,SONG HM,et al. miR-143-3p targeting LIM domain kinase 1 suppresses the progression of triple-negative breast cancer cells[J]. Am J Transl Res,2017,9(5):2276
[5] HE ZY,YI J,LIU XL,et al. MiR-143-3p functions as a tumor suppressor by regulating cell proliferation,invasion and epithelial-mesenchymal transition by targeting QKI-5in esophageal squamous cell carcinoma[J]. Mol Cancer,2016,15(1):51
[6]李勇,齐巍,王保苍,等. miR-143过表达对骨肉瘤143B细胞侵袭和迁移能力的影响[J].广东医学,2017,38(12):1809
[7] SHI HJ,SHEN HM,XU J,et al. MiR-143-3p suppresses the progression of ovarian cancer[J]. Am J Transl Res,2018,10(3):866
[8] YE Z,JINGZHONG L,YANGBO L,et al. Propofol inhibits proliferation and invasion of osteosarcoma cells by regulation of microRNA-143 expression[J]. Oncol Res,2013,21(4):201
[9] YANG C,GAO J,YAN N,et al. Propofol inhibits the growth and survival of gastric cancer cells in vitro through the upregulation of ING3[J]. Oncol Rep,2017,37(1):587
[10]DU Q,LIU J,ZHANG XE,et al. Propofol inhibits proliferation,migration,and invasion but promotes apoptosis by regulation of Sox4 in endometrial cancer cells[J]. Braz J Med and Biol Res,2018,51(4):e6803
[11]SUN H,GAO DY. Propofol suppresses growth,migration and invasion of A549 cells by down-regulation of miR-372[J]. BMC Cancer,2018,18(1):1252
[12]甘麦邻,杨露,谭娅,等. miR-143生物学功能的研究进展[J].生物技术通报,2017,33(6):16
[13]ZHAI L,MA C,LI W,et al. miR-143 suppresses epithelialmesenchymal transition and inhibits tumor growth of breast cancer through down-regulation of ERK5[J]. Mol Carcinog,2015,55(12):1990
[14]LEI C,DU F,SUN LA,et al. miR-143 and miR-145 inhibit gastric cancer cell migration and metastasis by suppressing MYO6[J]. Cell Death Dis,2017,8(10):e3101
[15]WANG L,HE J,XU H,et al. MiR-143 targets CTGF and exerts tumor-suppressing functions in epithelial ovarian cancer[J]. Am J Transl Res,2016,8(6):2716
[16]HUANG X,TENG Y,YANG H,et al. Propofol inhibits invasion and growth of ovarian cancer cells via regulating miR-9/NF-kappa B signal[J]. Braz J Med and Biol Res,2016,49(12):e5717