同源异型盒基因HOXA5对胃癌细胞BGC823侵袭能力的影响

Effect of HOXA5 on the Invasion Ability of Human Gastric Cancer Cell Line BGC823

目的检测人胃癌组织中HOXA5基因的表达情况,观察干扰HOXA5基因的表达之后,胃癌细胞BGC823的迁移和侵袭能力的变化,为阐明HOXA5在胃癌向远处转移的分子机制中提供新的思路。方法通过实时定量PCR(qRT-PCR)、Western blot以及免疫组织化学等技术,检测胃癌组织中HOXA5基因的表达。应用细胞转染的方法将HOXA5基因的过表达质粒转入胃癌细胞BGC823中,经qRT-PCR和Western blot验证HOXA5表达上调之后,利用Transwell侵袭实验检测BGC823细胞侵袭能力的变化。结果在收集的69例新鲜的胃癌组织以及相对应的癌旁组织中,免疫组织化学显示相比于癌旁组织的表达情况,胃癌组织中只有26例HOXA5的表达高于癌旁组织,比例为37.7%;qRTPCR、Western blot检测发现相对于癌旁组织,胃癌组织中HOXA5的表达水平显著下调(mRNA水平下调3倍,蛋白水平下调2倍,差异具有统计学意义,P<0.05)。BGC823细胞转染了过表达质粒之后,HOXA5的表达能够被上调。过表达了HOXA5基因的BGC823细胞的侵袭能力下降。结论在胃癌组织中HOXA5的表达下调,而在胃癌细胞BGC823中增强HOXA5的表达能够降低细胞的侵袭能力,说明HOXA5在胃癌的发生发展和侵袭、转移中发挥着抑癌基因的作用,可作为一个新的治疗靶点。

Objective To detect the expression of HOXA5 in human gastric carcinoma; and observe the changes of gastric cancer cell BGC823＇s migration and invasion ability after interference the expression of HOXA5 gene,and provide a new idea to clarify the molecular mechanism role of HOXA5 in the metastasis of gastric cancer. Methods Real-time quantitative PCR（ qRT-PCR）,Western blot and immunohistochemistry were performed to detect the expression of HOXA5 gene in gastric cancer tissues. The overexpression plasmid of HOXA5 was transferred into gastric cancer cell BGC823 by the method of cell transfection,we use qRT-PCR and western blot to confirm the overexpression effect,and then the changes of invasion ability of BGC823 cells were detected by transwell invasion assay. Results In the 69 cases of fresh gastric cancer tissue and the corresponding adjacent non-tumourous tissue,immunohistochemistry showed that in all gastric carcinoma tissue only 26 cases present HOXA5＇s expression was higher than that in adjacent non-tumourous tissue,the proportion was 37. 7%. Detection of qRT-PCR and western blot showed that the expression of HOXA5 was significantly lower in gastric cancer tissues than in adjacent non-tumourous tissue（ the level of mRNA was down 3 times,and the protein was 2 times,the difference was statistically significant,P〈0. 05）. In BGC823 cells,which have transfected overexpression plasmid of HOXA5,the expression of HOXA5 was up-regulated,and the invasion ability of BGC823 cells was decreased. Conclusion The expression of HOXA5 in gastric carcinoma decreased. Enhance the expression of HOXA5 in gastric cancer cell line BGC823 can reduce the invasion ability of cells. HOXA5 is a tumor suppressor in the development,invasion and metastasis of gastric cancer. It can be a new therapeutic target.