miR-628-3p靶向XPC调控皮肤癌细胞SK-MEL-5凋亡的机制

The Mechanism of mir-628-3p Targeting XPC to Regulate sk-mel-5 Apoptosis in Skin Cancer Cells

目的探讨miR-628-3p调控皮肤癌细胞SK-MEL-5的分子机制。方法使用实时荧光定量PCR检测人皮肤癌细胞SK-MEL-5、A2058、A-431和人正常皮肤细胞TE 353.Sk中的miR-628-3p表达量。miRDB在线分析miR-628-3p的潜在底物,并使用荧光素酶报告系统检测miR-628-3p与底物的结合。过表达或者敲低miR-628-3p后,通过Annexin V染色检测人皮肤癌细胞SK-MEL-5的凋亡水平。结果miR-628-3p在人皮肤癌细胞SK-MEL-5、A2058、A-431中的表达量均高于人正常皮肤细胞TE 353.Sk(P<0.05)。miR-628-3p靶向XPC的3端非编码区的708位置(P<0.05)。过表达miR-628-3p后,XPC的表达量下降,皮肤癌细胞SK-MEL-5的凋亡水平下降(P<0.05);敲低miR-628-3p后,XPC的表达量上升,皮肤癌细胞SK-MEL-5的凋亡水平上升(P<0.05)。结论miR-628-3p通过靶向XPC的3端非编码区来抑制皮肤癌细胞SK-MEL-5的凋亡。

Objective To investigate the molecular mechanism of mir-628-3p regulating sk-mel-5 in skin cancer cells.Methods The expressions of mir-628-3p in human skin cancer cells sk-mel-5,A2058,a-431 and human normal skin cells TE353.sk were detected by real-time fluorescence quantitative PCR.The potential substrates of mir-628-3p were analyzed online by miRDB and the binding of mir-628-3p to the substrates was detected by luciferase reporting system.After overexpression or knockdown of mir-628-3p,Annexin V staining was used to detect the apoptosis level of human skin cancer cell sk-mel-5,A2058 and a-431.Results The expression level of mir-628-3p in human skin cancer cell sk-mel-5,A2058 and a-431 was higher than that of human normal skin cell TE 353.sk(P<0.05).Mir-628-3 p targeted the 708 position of the 3-terminal non-coding region of XPC(P<0.05).After over-expressing mir-628-3p,the expression of XPC decreased and the apoptosis level of sk-mel-5 in skin cancer cells decreased(P<0.05).After knockdown of mir-628-3p,the expression of XPC increased and the apoptosis level of sk-mel-5 in skin cancer cells increased(P<0.05).Conclusion mir-628-3p inhibits the apoptosis of sk-mel-5 in skin cancer cells by targeting the 3-terminal non-coding region of XPC.