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目的婆罗双树样基因2(sal-like gene 2,SALL2)作为肿瘤抑制基因,在多种肿瘤细胞的生长过程中发挥调控作用。本研究旨在探讨SALL2基因对卵巢癌(ovarian cancer,OC)A2780细胞生长和转移的影响。方法采用小干扰RNA(small-interfering ribonucleic acid,siRNA)转染OC A2780细胞以沉默SALL2,设载体组及空白对照组。采用RT-PCR和蛋白质印迹法检测SALL2的表达。采用CCK-8和流式细胞术(flow cytometry,FCM)检测细胞增殖,采用FCM检测细胞凋亡。应用划痕实验检测细胞迁移,采用侵袭实验检测细胞侵袭。结果 A2780细胞高表达SALL2的mRNA及蛋白。RT-PCR结果显示,转染siRNA2和siRNA3 48h后,空白对照组、载体组、siRNA2和siRNA3组平均2-ΔΔCt值分别为1.000±0.030、0.942±0.053、0.207±0.041和0.465±0.007,差异有统计学意义,F=8.213,P=0.024。转染siRNA2和siRNA3 48h后,空白对照组、载体组及siRNA2和siRNA3组蛋白表达相对灰度比值分别为0.629±0.035、0.603±0.072、0.225±0.004和0.453±0.061,差异有统计学意义,F=11.629,P=0.018。siRNA2组转染72h后的细胞增殖力(4.285±0.026)显著高于载体组(3.622±0.029)和空白对照组(3.614±0.016),差异有统计学意义,F=34.023,P=0.012。siRNA2组转染48h后的细胞凋亡率为(49.17±2.03)%,显著低于载体组的(56.82±2.74)%和空白对照组的(58.39±2.45)%,差异有统计学意义,F=5.781,P=0.037。siRNA2组转染48h后处于G0/G1期的细胞比例为(47.87±1.28)%,均显著低于载体组的(55.27±1.46)%与空白对照组的(56.60±1.57)%,差异有统计学意义,F=4.213,P=0.032。siRNA2组转染48h后划痕愈合率为(78.925±6.133)%,明显高于载体组的(38.504±3.772)%和空白对照组的(42.169±4.103)%,差异有统计学意义,F=17.184,P=0.006。siRNA2组转染48h后穿过基质胶的细胞数(147.169±7.208)显著高于载体组(92.169±11.052)和空白对照组(95.169±9.830),差异有统计学意义,F=12.698,P=0.014。结论 SALL2沉默促进A2780细胞增殖、迁移和侵袭,抑制细胞凋亡。
Objective Sal-like gene 2 (SALL2) acts as a tumor suppressor gene and plays a regulatory role in the growth of various tumor cells. This study aimed to investigate the effect of SALL2 gene on the growth and metastasis of ovarian cancer (A2780) cells. Methods OC A2780 cells were transfected with small interfering ribonucleic acid (siRNA) to silence SALL2. The vector group and blank control group were established. SALL2 expression was detected by RT-PCR and Western blotting. Cell proliferation was detected by CCK-8 and flow cytometry (FCM), and apoptosis was detected by FCM. The scratch test was used to detect cell migration, and invasion assay was used to detect cell invasion. Results A2780 cells highly expressed SALL2 mRNA and protein. The results of RT-PCR showed that the average 2-ΔΔCt values of blank control group, vector group, siRNA2 and siRNA3 group were 1.000 ± 0.030,0.942 ± 0.053,0.207 ± 0.041 and 0.465 ± 0.007 respectively after transfected with siRNA2 and siRNA3 for 48h, the difference was Statistical significance, F = 8.213, P = 0.024. The relative gray value ratios of blank control group, vector group, siRNA2 and siRNA3 group were 0.629 ± 0.035,0.603 ± 0.072,0.225 ± 0.004 and 0.453 ± 0.061 respectively after transfection with siRNA2 and siRNA3 for 48h, the difference was statistically significant, F = 11.629, P = 0.018. The cell proliferation (4.285 ± 0.026) in siRNA2 group was significantly higher than that in vector group (3.622 ± 0.029) and blank control group (3.614 ± 0.016) at 72h after transfection. The difference was statistically significant (F = 34.023, P = 0.012). The apoptosis rate of siRNA2 group was (49.17 ± 2.03)% 48 h after transfection, which was significantly lower than that of vector group (56.82 ± 2.74)% and that of blank control group (58.39 ± 2.45)%, the difference was statistically significant = 5.781, P = 0.037. The percentage of cells in G0 / G1 phase of siRNA2 group was (47.87 ± 1.28)% at 48h after transfection, which were significantly lower than that of vector group (55.27 ± 1.46)% and that of blank control group (56.60 ± 1.57)% Significance, F = 4.213, P = 0.032. The wound healing rate in the siRNA2 group was (78.925 ± 6.133)% 48h after transfection, which was significantly higher than that in the control group (38.504 ± 3.772)% and the blank control group (42.169 ± 4.103)%, the difference was statistically significant, F = 17.184, P = 0.006. The number of cells passing through the matrigel in the siRNA2 group (147.169 ± 7.208) 48h after transfection was significantly higher than that in the vector group (92.169 ± 11.052) and the blank control group (95.169 ± 9.830), with a significant difference (F = 12.698, P = 0.014. Conclusion SALL2 silencing can promote proliferation, migration and invasion of A2780 cells and inhibit apoptosis.