摘要
目的探讨RGS22对人胃癌SGC-7901细胞在裸鼠体内转移的影响。方法将人RGS22基因克隆到逆转录病毒表达质粒LZRsPBMN中,构建重组质粒LZR-RGS22。以GFP为对照构建LZR-GFP。转染人胃癌SGC-7901细胞,Western blot法检测不同细胞中RGS22的表达。设计不同针对RGS22的RNAi片段和对照RNAi,将其构建入pSilenc-er2.1-U6 neo siRNA载体中,转染SGC-7901-LZR-RGS22细胞。RT-PCR法检测不同细胞中RGS22的表达。将不同组细胞注射于裸鼠腹腔内,观察不同肿瘤细胞的转移情况。结果 SGC-7901-LZR-RGS22细胞中146、112和76 ku片段RGS22蛋白表达较对照SGC-7901-LZR-GFP细胞表达均明显提高。RNAi 484片段和3313片段对RGS22的表达均有抑制作用。在裸鼠肿瘤转移实验中,RGS22能抑制SGC-7901肿瘤细胞向远处器官的转移,当其表达下调后,这种抑制效应又变弱。结论 RGS22能够抑制人胃癌SGC-7901在裸鼠中的转移。
Objective To study the role of RGS22 in SGC-7901 human gastric cancer cells on nude mice metastasis. Methods Human RGS22 gene was amplified by PCR, using eDNA library as template. Then it was inserted into retroviral expression vector LZRsPBMN, forming the recombinant plasmid LZR-RGS22. GFP, as a control, was constructed into LZR-GFP. I_ZR-RGS22 as well as LZR-GFP were transfected into SGC-7901 human gastric cancer cells. Western blot validated RGS22 expression in different cells. Different RNAi fragments for RGS22 and control RNAi were built into the pSilencer2. 1-U6 neo siRNA vector and transfected into SGC-7901-LZR-RGS22 cells. Real-time PCR detection of RGS22 expression in different cells. Different cell groups were injected intra-per- itoneally in nude mice to observe the tumor cell metastasis. Results Compared with the control SGC-7901-LZR - GFP cells, SGC-7901-LZR-RGS22 cells expressed higher 146 ku, 112 ku and 76 ku fragment of RGS22 protein. Compared with the control group, RNAi 484 and 3313 fragments inhibited RGS22 expression 24. 7% ± 7.7% and 13.8% ±2. 7% respectively. RGS22 could inhibit SGC-7901 tumor cells distant organs metastasis in nude mice experiments, while the inhibitory effect might be reduced when its expression was decreased. Conclusion RGS22 can inhibit SGC-7901 cell metastasis in nude mice.
出处
《安徽医科大学学报》
CAS
北大核心
2012年第6期650-654,共5页
Acta Universitatis Medicinalis Anhui
基金
江苏省教育厅创新基金(编号:JX22013009)
国家自然科学基金(编号:81000272,31071020)
