沉默RORα通过Wnt信号通路促进人胃癌细胞增殖与迁移侵袭
| 作者: |
1刘芳,
1,2赵晓红,
1夏红,
1,3苏坚,
1曾希,
1苏波,
1凌晖,
1苏琦
1 南华大学肿瘤研究所，湖南省胃癌研究中心，湖南省高校肿瘤细胞与分子病理学重点实验室，湖南 衡阳 421001 2 海南省妇幼保健院妇产科，海口 570206 3 南华大学附属第二医院，湖南 衡阳 421001 |
| 通讯: |
苏琦
Email: suqi1945@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2016.10.019 |
| 基金: | 国家自然基金, 81374013 湖南省卫计委科研课题, B2015-182 |
摘要
目的：在构建沉默RORα人胃癌MGC803细胞的基础上，观察沉默RORα对MGC803细胞增殖与迁移侵袭的影响。方法：软琼脂集落形成实验、流式细胞术、细胞迁移和侵袭实验检测RORα沉默对MGC803细胞增殖、细胞周期、迁移和侵袭能力的影响。RT-PCR与Western blot检测RORα、Wnt1、MMP-9与TIMP3 mRNA与蛋白表达。结果：集落形成实验显示，沉默组的集落形成率128.1%明显高于对照组和空载体组(P<0.05)。流式细胞术显示，沉默组S期细胞百分率39.9%较对照组26.1%和空载体组27.4%明显增加(P<0.05)。迁移实验显示，沉默组迁移距离(1.76±0.19) mm明显高于对照组(1.32±0.14) mm和空载体组(1.29±0.17) mm (P<0.05)。侵袭实验显示，沉默组穿膜细胞(172±27)个明显多于对照组(147±17)个和空载体组(149±14)个(P<0.05)。RT-PCR与Western blot表明，沉默RORα可显著上调Wnt1与MMP-9 mRNA与蛋白与下调RORα与TIMP3 mRNA与蛋白。结论：沉默RORα可通过Wnt信号通路上调MMP-9和下调TIMP3促进MGC803细胞增殖与迁移侵袭。
关键词：
RORα沉默
人胃癌MGC803细胞
增殖
迁移
侵袭
Wnt通路
Silencing RORα promote proliferation, migration and invasion through Wnt signaling pathway in MGC803 cells
CorrespondingAuthor: SU Qi Email: suqi1945@163.com
DOI: 10.3978/j.issn.2095-6959.2016.10.019
Abstract
Objective: To investigate Silencing RORα through Wnt signaling pathway promote proliferation, migration and invasion in human gastric MGC803 cells underlaying construction of MGC803 cells of silencing RORα. Methods: Colony formation and flow cytometry were used for cell proliferation and cell cycle. Wound healing and transwell assays were performed to examine cell migration and invasion activities. The expression of RORα, Wnt1, MMP-9 and TIMP3 mRNA and protein were detected by Western blot and RT-PCR. Results: The colony forming showed that the colony forming efficiency in miR-RORα group was 128.1% higher significantly than in control and vector/MGC803 (P<0.05). The flow cytometry revealed that percentage in S cycle was 39.9% in miR-RORα markedly increase than 26.1% in control and 27.4% in vector/MGC803 (P<0.05). The migration experiment disclosed that migration distance in miR-RORα was (1.76±0.19) mm notably increase than (1.32±0.14) mm in control and (1.29±0.17) mm in vector/MGC803 (P<0.05). The invasion essay exhibited that the cells of passed through the Matrigel were (172±27) in miR-RORα increased than (147±17) in control and (149±14) in vector/MGC803, respectively (P<0.05). RT-PCR and Western blot indicated that silencing RORα up-regulated expression of wnt1 and MMP-9 and down-regulated expression of RORα and TIMP3, compared with control and vector/MGC803, respectively (P<0.05). Conclusion: Silencing RORα can promote proliferation, migration and invasion through Wnt signaling pathway in human gastric MGC803 cells.