干扰Hela细胞TPX2基因对细胞活性及其放疗敏感性的影响研究

Effect of Interference of TPX2 Gene on Cell Activity and Radiotherapy Sensitivity of Hela Cells

目的:分析干扰Hela细胞TPX2基因对细胞的活性及放疗敏感性的影响。方法:体外培养宫颈癌Hela细胞系,并将其分为干扰组、阴性对照组(NC组)、空白对照组(CON组)。干扰组接受TPX2-siRNA干扰质粒、NC组接受空载体转染,CON组以PBS代替质粒,采用RT-PCR实验验证干扰效率,采用CCK-8实验检测细胞增殖能力,采用Transwell小室检测侵袭能力,流式细胞仪检测细胞周期,克隆形成实验检测放疗敏感性。结果:与CON组相比,NC组细胞的TPX2 mRNA相对表达量、TPX2蛋白相对表达水平、细胞侵袭能力均无明显变化(P>0.05);与CON组及NC组相比,干扰组细胞的TPX2 mRNA相对表达量、TPX2蛋白相对表达水平、细胞侵袭能力均明显降低(P<0.05)。与CON组相比,NC组细胞的细胞活性在各个时间点差异均无统计学意义(P>0.05);与CON组及NC组相比,干扰组细胞在培养时间为48h、72h、96h时的细胞活性均明显降低(P<0.05)。与CON组相比,NC组的G0/G1期、G2/M期及S期的细胞周期占比无明显改变(P>0.05);与NC组相比,干扰组细胞的S及G2/M期细胞占比明显增多,而G0/G1期细胞占比明显减少(均P<0.05)。与CON组相比,干扰组细胞在放疗剂量为4Gy、6Gy、8Gy、10Gy的生存率降低(P均<0.05)。结论:干扰宫颈癌Hela细胞TPX2基因可有效干扰TPX2基因的表达,阻碍宫颈癌Hela细胞的增殖、迁移等细胞活性。干扰TPX2基因可有效提高宫颈癌Hela细胞的放疗敏感性,改善宫颈癌放射治疗的疗效。

Objective:To analyze the effects of interference with TPX2 gene on the activity and radiotherapy sensitivity of HeLa cells.Methods:Cervical cancer Hela cell is cultured in vitro and divided into interference group,negative control group(NC group),blank control group(CON group),interference and CON group.Respectively the interference group had TPX2-siRNA interference plasmid;CON group had plasmid transfection,replacing the plasmid with PBS;NC group had blank load transfection.CCK 8 experimental detection of efficiency of cell proliferation,Transwell Chambers detection of invasion ability,clone forming experiments to detect radiation sensitivity were all conducted.Results:Compared with the CON group,the relative expression level of TPX2 mRNA,the relative expression level of TPX2 protein,and the cell invasion ability of the NC group did not change significantly(P>0.05);compared with the CON group and the NC group,the TPX2 of the interference group,the relative expression of mRNA,the relative expression of TPX2 protein,and the ability of cell invasion were all significantly reduced(P<0.05).Compared with the CON group,the cell viability of the NC group was not significantly different at each time point(P>0.05);compared with the CON group and the NC group,the culture time of the interference group was 48h,72h,96h.The cell viability at time was significantly reduced(P<0.05).Compared with the CON group,the proportion of cell cycles in the G0/G1,G2/M and S phases of the NC group did not change significantly(P>0.05);compared with the NC group,the S and G2/The proportion of cells in M phase increased significantly,while the proportion of cells in G0/G1 phase decreased significantly(P<0.05).Compared with the CON group,the survival rate of the interference group cells at 4Gy,6Gy,8Gy,and 10Gy doses of radiotherapy decreased(P<0.05).Conclusion:Interference with TPX2 gene in cervical cancer Hela cells can effectively interfere with the expression of TPX2 gene and inhibit the cell activity of proliferation and migration of cervical cancer HeLa cells.Interference with TPX2 gene can effectively increase the radiotherapy sensitivity of cervical cancer HeLa cells and improve the curative effect of radiotherapy for cervical cancer.