PKC信号通路在KISS1基因抑制人结直肠癌HCT116细胞侵袭迁移的作用

发布时间：2018-06-22 19:52

  本文选题：结直肠癌 + KISS基因　； 参考：《重庆医科大学学报》2017年11期

【摘要】：目的:探讨PKC信号通路在KISS1基因发挥抑制结直肠癌细胞HCT116侵袭迁移的作用。方法:构建p GC-LV-KISS1-EGFP慢病毒载体感染人结直肠癌细胞HCT116,分空白对照组(CON组)、空载体对照组(NC组)、过表达组(OE组)。q RT-PCR和Western blot检测KISS1基因m RNA和Metastin、PKC信号通路的关键分子PKCα、PKCβII及下游E钙黏蛋白表达量的变化。Transwell法检测细胞侵袭、迁移能力的变化。结果:转染后能够稳定表达报告基因EGFP,荧光率均80%,且OE组KISS1基因m RNA、Metastin表达量较CON组和NC组均明显升高(P0.05)。转染后,PKCβⅡ的蛋白表达量(0.288 2±0.023 7)较CON组(0.530 9±0.013 3)和NC组(0.511 7±0.008 5)明显下降(P0.05),而PKCα的表达水平无明显变化(P0.05);下游效益蛋白E钙黏蛋白表达量(0.633 2±0.017 4)较CON组(0.232 2±0.019 6)和NC组(0.252 3±0.018 2)明显升高(P0.05)。OE组细胞侵袭、迁移能力较CON组和NC组均明显下降(P0.05)。结论:KISS1基因可能通过下调PKCβⅡ后上调下游蛋白E钙黏蛋白的表达而发挥抑制结直肠癌细胞HCT116侵袭、迁移作用,有望成为防治结直肠癌转移的新靶点。
[Abstract]:Aim: to investigate the role of PKC signaling pathway in inhibiting invasion and migration of colorectal cancer cell line HCT116. Methods: HCT116 cells infected with pGC-LV-KISS1-EGFP lentivirus vector were divided into blank control group (Con group), empty vector control group (NC group), overexpression group (OE group). Q RT-PCR and Western blot were used to detect KISS1 gene mRNA and PKC 伪 PKC 尾 II signal pathway. The expression of E-cadherin was detected by Transwell method. A change in mobility. Results: EGFP could be expressed stably after transfection, the fluorescence rate was 80%, and the expression of KISS1 gene m RNA-Metastin in OE group was significantly higher than that in Con group and NC group (P0.05). After transfection, the protein expression of PKC 尾 鈪，

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