Sestrin2在Eupatilin调控结直肠癌细胞株SW480增殖和凋亡中的作用研究

Study on the role of Sestrin2 in Eupatilin regulating the proliferation and apoptosis of colorectal cancer SW480 cell line

目的探讨Sestrin2(SESN2)在Eupatilin抑制结直肠癌细胞株SW480增殖中的作用。方法分别以短发夹RNA慢病毒稳定抑制SESN2表达载体(shSESN2)及其对照乱序非编码片段(shCon)感染SW480细胞以制备SW480shSESN2细胞和SW480^shCon细胞。以SW480^shCon和SW480shSESN2细胞为研究对象,CCK-8法检细胞增殖能力并计算Eupatilin对各细胞的半数抑制浓度(IC50)。以Eupatilin对SW480^shCon细胞的IC50浓度(37.93μmol/L)干预两组细胞48 h后,流式细胞术检测细胞凋亡,蛋白印迹检测细胞Sestrin2、LC3、P62等蛋白的表达。结果与SW480^shCon细胞相比,SW480shSESN2细胞活力增高、Eupatilin对其的IC50增加。经Eupatilin干预48 h后,与SW480shSESN2细胞相比,SW480^shCon细胞凋亡增加,P62蛋白表达降低、LC3Ⅱ/Ⅰ蛋白表达比例升高。结论Eupatilin可能通过激活Sestrin2以抑制结肠癌细胞株SW480增殖、促进其凋亡。

Objective To investigate the role of Sestrin2 in Eupatilin regulating the proliferation and apoptosis of colorectal cancer SW480 cell line.Methods Lentivirus(LV)-mediated short hairpin RNA(shRNA)targeting to inhibit sestrin2(shSESN2)expression and the controlled non-silent shRNA(shCon)vector were used to transfected the SW480 cells to engineer SW480shSESN2 cells and SW480^shCon cells.The different final concentrations(0 to 100μmol/L)of Eupatilin were used to coculture with SW480^shConand SW480shSESN2 cells for 48 h.Cell counting kit-8(cck-8)was used to detect the proliferation of the cells and calculate the half inhibitory concentration(IC50)of Eupatilin for each group.Eupatilin with IC50 of SW480 cells(37.93μmol/L)was used to intervene the cells for 48 h.Cells apoptosis was detected by flow cytometry,and the expression of Sestrin2,LC3 and P62 proteins in cells was detected by western blot.Results Compared with SW480^shCon cells,SW480shSESN2 cells showed increased proliferation and IC50 of Eupatilin,decreased apoptosis rate,increased protein expression of P62,and decreased protein expression ratio of LC3Ⅱ/Ⅰ.Conclusion Eupatilin may inhibit the proliferation and promote the apoptosis of colon cancer cell line SW480 by activating sestrin2.