龙胆苦苷通过调控LncRNA LINC00324/miR-3619-5p表达对皮肤鳞癌SCL-1细胞增殖、迁移及侵袭的影响

Effect of Gentiopicroside on Proliferation,Migration,and Invasion of Cutaneous SquamousCell Carcinoma SCL-1 Cells by Regulating the Expression of LncRNA LINC00324/miR-3619-5p

目的探讨龙胆苦苷(Gentiopicroside,GPS)对皮肤鳞癌SCL-1细胞增殖、迁移、侵袭的影响及其对LncRNA LINC00324/miR-3619-5p的调控作用。方法采用不同浓度的GPS处理人皮肤鳞癌细胞SCL-1(GPS-L组、GPS-M组、GPS-H组),同时将正常培养的细胞记为Con组,si-NC、si-LINC00324分别转染至si-NC组、si-LINC00324组,pcDNA、pcDNA-LINC00324分别转染入SCL-1细胞后加入40μmol/L GPS处理细胞(GPS+pcDNA组、GPS+pcDNA-LINC00324组);采用MTT实验、Transwell小室实验分别检测细胞增殖、迁移及侵袭能力;采用qRT-PCR法检测LINC00324、miR-3619-5p的表达量;双荧光素酶报告实验检测LINC00324与miR-3619-5p的靶向关系。结果与Con组比较,GPS-L组、GPS-M组、GPS-H组细胞增殖抑制率升高(P<0.05),细胞转移及侵袭数量明显降低(P<0.05),LINC00324的表达水平降低(P<0.05),miR-3619-5p的表达水平升高(P<0.05);双荧光素酶报告实验证实LINC00324可靶向结合miR-3619-5p;与si-NC组比较,si-LINC00324组细胞增殖抑制率升高(P<0.05),迁移及侵袭细胞数减少(P<0.05);转染pcDNA-LINC00324可减弱GPS对SCL-1细胞增殖、迁移及侵袭的作用。结论龙胆苦苷可通过抑制LINC00324的表达而促进miR-3619-5p的表达,进一步降低皮肤鳞癌SCL-1细胞增殖、迁移及侵袭的能力。

Objective To explore the effect of gentiopicroside(GPS)on the proliferation,migration,and invasion of cutaneous squamouscell carcinoma SCL-1 cells and its regulation onLncRNA LINC00324/miR-3619-5 p.Methods Different concentration of GPS was used to treat human cutaneous squamouscell carcinoma SCL-1 cells(GPS-L group,GPS-M group,and GPS-H group),and the normally cultured cells were recorded as the Con group.Thesi-NC and si-LINC00324 were transfected into the si-NC group and the si-LINC00324 group,respectively.The pc DNA and pcDNA-LINC00324 were transfected into SCL-1 cells,and then 40μmol/L GPStreated cells were added(GPS+pcDNA group and GPS+pcDNA-LINC00324 group).The methylthiazolyl tetrazolium(MTT)experiment and Transwell chamber experiment were used to detect the cell proliferation,migration,and invasion ability,respectively.The real-time quantitative polymerase chain reaction(qRT-PCR)method was used to detect the expressions of LINC00324 and miR-3619-5 p.The dual luciferase reporter experiment was used to detect the targeting relationship between LINC00324 and miR-3619-5 p.Results As compared with the Con group,the cell proliferation inhibition rate in the GPS-L group,GPS-M group,and GPS-H group was increased(P<0.05),and the number of migration and invasion cells was decreased(P<0.05).The expression level of LINC00324 was decreased(P<0.05),and the expression level of miR-3619-5 p was increased(P<0.05)in these groups.The dual luciferase report experiment confirmed that LINC00324 targeted to miR-3619-5 p.As compared with the si-NC group,the cell proliferation inhibition rate in the si-LINC00324 group was increased(P<0.05),and the number of migration and invasion cells was reduced(P<0.05).Transfection of pcDNA-LINC00324 attenuated the effect of GPS on the proliferation,migration,and invasion of SCL-1 cells.Conclusion GPS promotes the expression of miR-3619-5 p by inhibiting the expression of LINC00324,thereby reducing the ability of cutaneous squamouscell carcinoma SCL-1 cells to proliferate,migrate,and invade.