摘要
目的:探讨SEMA4C在胃癌中的表达以及hsa-miR-19a-3p下调SEMA4C对胃癌细胞增殖、侵袭能力的影响。方法:收集23例胃癌组织和对应癌旁组织标本,采用免疫组化染色、Real-time PCR方法检测SEMA4C的表达,并分析其与临床资料的相关性;检索Targetscan、Miranda等数据库预测调控SEMA4C基因的microRNA,并利用Real-time PCR、双荧光素酶报告系统检测加以证实;通过脂质体转染使人胃癌细胞系BGC823内过表达hsa-miR-19a-3p,Real-time PCR、Western blot检测SEMA4C的表达,通过细胞集落形成实验和Transwell小室研究BGC823细胞的增殖、侵袭能力的变化。结果:胃癌组织中SEMA4C阳性率87.0%,高于癌旁组织(χ2=20.30,P<0.000 1),胃癌组织中SEMA4 C mRNA的表达高于癌旁组织(t=13.47,P<0.000 1)。SEMA4C的表达与胃癌TNM分期(P=0.036 4)、分化程度(P=0.042 7)、淋巴结转移度(P=0.012 8)有关。生物信息学分析hsa-miR-19a-3p、hsa-miR-214-5p、hsa-miR-138-5p可能调控SEMA4C表达,胃癌组织中hsa-miR-19a-3p的表达低于癌旁组织(t=8.233,P<0.000 1),hsamiR-214-5p(t=0.846 3,P=0.097 6)、hsa-miR-138-5p(t=1.345,P=0.185 7)表达与癌旁组织无明显差异,hsa-miR-19a-3p转染后荧光素酶表达下降至0.46±0.12(F=5.685,P=0.003 2)。hsa-miR-19a-3p转染后,SEMA4C mRNA(F=34.39,P<0.000 1)、SEMA4C蛋白(F=67.49,P<0.000 1)表达减少,BGC823细胞克隆数目减少(F=20.77,P<0.000 1),Transwell下室细胞数较少(F=16.37,P=0.000 4)。结论:SEMA4C在胃癌中过表达并与肿瘤的发生发展有关,hsa-miR-19a-3p通过下调靶基因SEMA4C抑制胃癌细胞增殖、侵袭能力来发挥其抗肿瘤效应。
Objective：To investigate the expression of SEMA4C in gastric cancer and the effect of hsa-miR-19a-3p on proliferation and invasion of gastric cancer cell by down-regulation of SEMA4C.Methods：The expression of SEMA4C was determined in 23 patients with gastric cancer and the relationship between SEMA4C and clinical features was analyzed.The probable microRNAs were predicted by TargetScan and Miranda,and tested by Real-time PCR and Dual luciferase reporter system.The expression of SEMA4C was determined after BGC823 cells were transfected with hsa-miR-19a-3p,and the effects of hsa-miR-19a-3p on proliferation and invasion of BGC823 cells were tested by Transwell assay.Results：The positive rate of SEMA4C was significantly higher in gastric tissues than that in para-cancer tissues （x2=20.30,P〈0.000 1）,and the level of SEMA4C mRNA was also higher in gastric tissues （t=13.47,P〈0.000 1）.The expression of SEMA4C was associated with TNM stage （P=0.036 4）,differentiation （P=0.042 7） and lymph nodes metastasis（P=0.012 8）.The expression of hsa-miR-19a-3p was lower in gastric tissues than that in para-cancer tissues（t=8.233,P〈0.000 1）.After hsa-miR-19a-3p was transfected to BGC823 cells,the expression of SEMA4C was decreased,and the proliferation and invasion of gastric cancer cell were inhibited.Conclusion：The expression of SEMA4C is down-regulated in gastric cancer tissues,and hsa-miR-19a-3p inhibits proliferation and invasion of gastric cancer cell by down-regulation of SEMA4C.
出处
《现代肿瘤医学》
CAS
2017年第16期2609-2615,共7页
Journal of Modern Oncology
基金
陕西省自然科学基础研究计划(编号:2015JM8389)
关键词
胃肿瘤
微RNAS
增殖
侵袭
gastricneoplasm
microRNAs
proliferation,invasion
