摘要
目的观察Rab1A在胃癌组织的表达及其对胃癌细胞SGC7901增殖和凋亡的影响。方法收集35例胃癌患者的胃癌组织及正常胃组织,用real-time PCR和Western blot法检测胃癌组织和正常胃组织中Rab1A表达水平。SGC7901胃癌细胞分为对照组,阴性对照组,Rab1A siRNA 30 nmol/L组,60 nmol/L组和90 nmol/L组,分别给予无效处理、阴性对照siRNA和三种不同浓度的Rab1A siRNA,MTT法检测各组细胞增殖活力。60 nmol/L的Rab1A siRNA沉默SGC7901细胞中Rab1A的表达,流式细胞仪分析Rab1A siRNA对细胞周期和细胞凋亡的影响,Western blot法检测Rab1A siRNA对Bax和Bcl-2蛋白表达的影响。结果胃癌组织中Rab1A的表达无论在mRNA水平(5.625±0.525 vs 1.000±0.000,P<0.05)还是蛋白水平(1.013±0.125 vs 0.219±0.060,P<0.05),均显著高于正常胃组织。RNA干扰48 h后,与对照组相比,30 nmol/LRab1A siRNA组的细胞增殖活力无明显变化(P>0.05),60 nmol/LRab1A siRNA组和90 nmol/LRab1A siRNA组的细胞增殖活力明显下降(P<0.05),但60 nmol/L组和90 nmol/L组间细胞增殖活力无明显差异(P>0.05)。与对照组相比,60 nmol/L siRNA组无论是G1/G0期、S期,还是G2/M期的细胞数量均无明显差异(P>0.05);与对照组相比,60 nmol/L siRNA组无论是早期凋亡细胞比例,还是晚期凋亡细胞的比例均明显上升(P<0.05);而且,与对照组相比,60 nmol/L siRNA组的Bax蛋白的表达水平显著上调(0.483±0.015 vs 0.767±0.153,P<0.05),Bcl-2蛋白的表达水平显著下调(0.703±0.021 vs 0.347±0.015,P<0.05)。结论 Rab1A在胃癌组织中的表达水平高于正常胃组织,可促进胃癌细胞SGC7901增殖,并且通过上调Bcl-2和下调Bax的表达抑制细胞凋亡。
Objective To explore the expression of Rab1 A in gastric cancer tissues and its effects on the proliferation and apoptosis of SGC7901 gastric cancer cells. Methods Gastric cancer tissues and normal gastric tissues from 35 gastric cancer patients were collected,and Rab1 A expression was analyzed by real-time PCR and Western blot. SGC7901 gastric cancer cells were divided into control group,negative control siRNA group,30 nmol / L siRNA group,60 nmol / L group and 90 nmol / L group. MTT assay was used to detect the cell proliferation abilities. After Rab1 A expression was silenced by 60 nmol / L Rab1 A siRNA,the cell cycle and apoptosis of SGC7901 gastric cancer cells were examined with flow cytometer,and the expression levels of Bax and Bcl-2 protein were analyzed with Western blot. Results Rab1 A expression was higher in human gastric cancer tissues than in normal gastric tissues both at the mRNA level（ 5. 625 ± 0. 525 vs 1. 000 ± 0. 000,P〈0. 05） and at the protein level（ 1. 013 ± 0. 125 vs 0. 219 ± 0. 060,P〈0. 05）. At 48 h after RNA interference,the cell proliferation ability showed no significant difference between 30 nmol / L Rab1 A siRNA group and control group（ P〈0. 05）,and the cell proliferation abilities in 60 nmol / L and 90 nmol / L Rab1 A siRNA groups decreased markedly compared with control group（ P〈0. 05）,but they showed no significant difference between 60 nmol / L group and 90 nmol / L group（ P〈0. 05）.The numbers of cells in G1/ G0,S and G2/ M phases showed no difference between 60 nmol / L Rab1 A siRNA group and control group.Both early apoptotic rate and late apoptotic rate in 60 nmol / L Rab1 A siRNA group decreased markedly compared with control group（ P〈0. 05）. Compared with control group,Bax protein expression increased（ 0. 483 ± 0. 015 vs 0. 767 ± 0. 153,P〈0. 05） and Bcl-2 protein expression decreased significantly（ 0. 703 ± 0. 021 vs 0. 347 ± 0. 015,P〈0. 05） in 60 nmol / L Rab1 A siRNA group. Conclusion Rab1 A is highly expressed in hum
出处
《山西医科大学学报》
CAS
2016年第10期895-901,共7页
Journal of Shanxi Medical University
基金
中国博士后科学基金面上资助项目(2013M542358)
陕西省自然科学基金资助项目(2014JM4122)
关键词
Rab1A
胃癌
细胞增殖
细胞凋亡
Rab1A
gastric cancer
cell proliferation
cell apoptosis
