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LncRNA ANCR介导PI3K及AKT蛋白对人胃癌细胞增殖 迁移的作用机制
朱金庚,刘永,涂志刚
0
(解放军中部战区总医院汉口院区普外科)
摘要:
【摘要】 目的 探讨LncRNAANCR介导PI3K及AKT蛋白对人胃癌细胞增殖、迁移的作用机制。方法 选取人胃癌细胞株,随机分为胃癌组(GC组)、ANCRsiRNA组(siRNA组)、LncRNAANCR组(ANCR组)3组,同时选取正常胃粘膜上皮细胞设为正常胃细胞组。GC组:单纯胃癌细胞株;siRNA组:将ANCRsiRNA转染到胃癌细胞中;ANCR组:将LncRNAANCR转染到胃癌细胞中。通过TUNEL法、Transwell、cck8法、Westernblot法、RTPCR法分析3组胃癌细胞增殖情况,PI3K、AKT蛋白表达量,细胞凋亡情况,细胞侵袭能力,PI3K、AKT mRNA的表达量的差异性来探究LncRNAANCR介导PI3K及AKT蛋白对人胃癌细胞增殖迁移的作用机制。结果 GC组LncRNAANCR蛋白含量最高,正常胃细胞组LncRNAANCR蛋白含量最低(P<0.05);ANCR组胃癌细胞侵袭能力强,显微镜下细胞数量多,siRNA组胃癌细胞显微镜下细胞数量少,侵袭能力弱(P<0.05),GC组与siRNA组相比较胃癌细胞数量明显增多,整体侵袭能力明显增强(P<0.05);GC组和siRNA组、ANCR组OD值相比较,差异均有统计学意义(P<0.05);ANCR组胃癌细胞的增殖情况明显高于GC组,而GC组明显高于siRNA组,差异均有统计学意义(P<0.05)。经TUNEL染色,siRNA组胃癌细胞凋亡数量最多,ANCR组胃癌细胞凋亡较少(P<005),ANCR组和GC组与siRNA组相比较凋亡率明显下降(P<0.05)。蛋白免疫印迹法分析显示,siRNA组的PI3K、AKT蛋白含量及表达量最低,ANCR组的PI3K、AKT蛋白含量及表达量最高,3组相比较差异均有统计学意义(P<0.05)。结论 LncRNAANCR能激活PI3K及AKT蛋白的表达,升高细胞的侵袭和增殖能力,促进胃癌细胞生长,加快病情的发展。
关键词:  胃癌; LncRNA ANCR;PI3K,AKT;细胞增殖  迁移
DOI:
基金项目:湖北省自然科学基金课题
LncRNA ANCR mediates the mechanism of PI3K and AKT protein on proliferation and migration of human gastric cancer cells
ZHU Jingeng,LIU Yong,TU Zhigang
(General Surgery Department, Hankou Hospital, Central War Zone General Hospital of PLA)
Abstract:
【Abstract】 Objective To investigate the mechanism of LncRNAANCR mediating the proliferation and migration of human gastric cancer cells by PI3K and AKT proteins.Methods Human gastric cancer cell lines were randomly divided into gastric cancer group (GC group), ANCR siRNA group (siRNA group) and LncRNA ANCR group (ANCR group). At the same time, normal gastric epithelial cells were selected as normal gastric cell group. GC group: simple gastric cancer cell line. siRNA group: ANCR siRNA was transfected into gastric cancer cells; ANCR group: LncRNA ANCR was transfected into gastric cancer cells. The proliferation, PI3K and AKT protein expression, apoptosis and invasive ability of gastric cancer cells in three groups were analyzed by TUNEL, Transwell, CCK 8, Western blot and RT-PCR. Differential expression of PI3K and AKT mRNA were explored the mechanism of LncRNA ANCR mediating PI3K and AKT protein on proliferation and migration of human gastric cancer cells.Results Transwell invasion experiments were performed on three groups of gastric cancer cells. It is obvious that the gastric cancer cells in the ANCR group had strong invasive ability, and the number of cells under the microscope is large. The number of cells in the siRNA group under the microscope was small, and the invasive ability is weak (P<0.05). The number of infections increased significantly, and the overall invasive ability was significantly enhanced (P<0.05). The OD values between the siRNA group and the GC group were significantly different between the two groups. The OD values of the GC group and the ANCR group were also significantly different. The proliferation of gastric cancer cells in the ANCR group was the highest, and the proliferation of the gastric cancer cells in the GC group was the least. The proliferation of gastric cancer cells in the ANCR group was significantly higher than that in the GC group. The proliferation of gastric cancer cells in the GC group was significantly higher than that in the siRNA group (P<0.05). After TUNEL staining, the apoptosis of gastric cancer cells was the highest in siRNA group, and the apoptosis of gastric cancer cells in ANCR group was less (P<0.05). The apoptosis rate of ANCR group was significantly lower than that of GC group and siRNA group (P<0.05). Western blotting was performed on GC group, siRNA group and ANCR group. The analysis showed that the siRNA group had the lowest content of PI3K and AKT protein, the ANCR group had the highest content of PI3K and AKT protein, and the ANCR group compared with GC group and siRNA group PI3K, AKT. Protein expression was significantly (all P<0.05). The results showed that the expression of PI3K and AKT mRNA was the lowest in gastric cancer cells of siRNA group, and the expression of PI3K and AKT mRNA was the highest in gastric cancer cells of ANCR group. The expression of PI3K and AKT mRNA in ANCR group was significantly higher than that in GC group and siRNA group (both P<0.05). Conclusion LncRNAANCR can activate the expression of PI3K and AKT protein, increase the invasion and proliferation of cells, promote the growth of gastric cancer cells, and accelerate the development of the disease.
Key words:  Gastric cancer  LncRNA ANCR  PI3K  AKT  Proliferation  Migration

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