婆罗双树样基因2对A2780细胞生长及转移影响

发布时间：2018-06-13 04:13

  本文选题：SALL + 卵巢癌　； 参考：《中华肿瘤防治杂志》2017年07期

【摘要】：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。
[Abstract]:Objective as a tumor suppressor gene, 2(sal-like gene _ 2 and SALL2) play a regulatory role in the growth of various tumor cells. The aim of this study was to investigate the effect of SALL2 gene on the growth and metastasis of ovarian cancer cell line OCCAN A2780. Methods OC-A2780 cells were transfected with small interfering ribonucleic siRNAs to silence SALL2. Vector group and blank control group were set up. The expression of SALL2 was detected by RT-PCR and Western blotting. CCK-8 and flow cytometric FCM were used to detect cell proliferation and FCM to detect cell apoptosis. Scratch test was used to detect cell migration and invasion test was used to detect cell invasion. Results after transfection of siRNA2 and siRNA3 for 348h, the average values of 2- 螖 Ct in control group, vector group and siRNA3 group were 1.000 卤0.030, 0.942 卤0.053, 0.207 卤0.041 and 0.465 卤0.007, respectively. The difference was statistically significant. After transfection of siRNA2 and siRNA3 for 48 h, the control group was treated with siRNA2 and siRNA3. The relative gray ratio of protein expression in vector group, siRNA2 and siRNA3 group was 0.629 卤0.035 卤0.072 卤0.225 卤0.004 and 0.453 卤0.061, respectively. The difference was statistically significant (P < 0.01). The proliferative power of the vector group was significantly higher than that of the vector group (3.622 卤0.026) and the blank control group (3.614 卤0.016), and the difference was statistically significant. After 48 h transfection, the cell apoptosis rate was 49.17 卤2.03g, which was significantly lower than that in the vector group (56.82 卤2.74%) and the blank control group (58.39 卤2.45%). The difference was statistically significant (P < 0.01). The percentage of cells in the G _ 0 / R _ 1 phase of the control group was 47.87 卤1.28%, which was significantly lower than that of the vector group (55.27 卤1.46g%) and the blank control group (P < 0.05). 56.60 卤1.57, the difference was statistically significant (P < 0.05). The rate of scratch healing in group 2 was 78.925 卤6.133 after transfection for 48 h, which was significantly higher than that in vector group (38.504 卤3.772%) and blank control group (42.169 卤4.103%). The difference was statistically significant. The number of cells passing through the matrix gel in group 2 was significantly higher than that in control group (147.169 卤7.208). The difference between the control group (92.169 卤11.052) and the blank control group (95.169 卤9.830) was statistically significant (P < 0.05). Conclusion SALL2 silencing can promote the proliferation, migration and invasion of A2780 cells and inhibit the apoptosis of A2780 cells.
【作者单位】： 滨州医学院免疫学教研室;
【基金】：国家自然科学基金(81001300)
【分类号】：R737.31
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本文编号：2012672