新基因SPATA12抑制肿瘤细胞增殖的实验研究  

Experimental study of inhibition of tumor cell proliferation by a novel gene SPATA12

作　　者：刘志文[1] 林奕婷[1] 刘选明[1] 余伟伟[1] 章运生[1] 李丹[1] 

机构地区：[1]湖南大学生物学院生命科学系,长沙410082

出　　处：《中南大学学报（医学版）》2012年第3期222-227,共6页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(30872763);中央高校基本科研业务费专项资金(531107040314);湖南省高校青年骨干教师资助项目(521298462)~~

摘　　要：目的:探讨新基因精子发生相关基因12(spermatogenesis associated gene 12,SPATA12)对肿瘤细胞增殖的抑制作用及可能机制。方法:采用组织芯片原位杂交技术分析SPATA12在睾丸肿瘤中的表达谱;四甲基偶氮唑盐[3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide,MTT]比色法、平板集落形成实验分析SPATA12对肿瘤细胞增殖能力、集落形成能力的影响;反转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)分析SPATA12对肿瘤细胞周期基因表达水平的改变。结果:组织芯片原位杂交结果发现SPATA12基因在正常睾丸组织表达,而在精原细胞瘤等睾丸肿瘤中均表达缺失;MTT和平板集落形成实验表明SPATA12可明显地抑制肿瘤细胞的增殖能力和集落形成能力;RT-PCR实验显示SPATA12可抑制周期蛋白(cyclin)A1的表达,使cyclin D1表达下调,而对cyclin B1和cyclin E1无显著影响。结论:SPATA12可能在肿瘤的发生中扮演抑制因子的角色,并通过调控细胞周期基因发挥作用。Objective： To explore the inhibitory role of spermatogenesis-associated gene 12 （SPATA12） on tumor cell proliferation and its possible mechanism. Methods： The expression pattern of SPATA12 in testicular tumors was investigated by in situ hybridization analysis using tissue microarrays. The effects of SPATA12 on tumor cell proliferation and colony formation was detected by 3-（4.5-dimethylthiazol-2- yl）-2,5-diphenyl tetrazolium bromide （MTF） assay and colonyforming assays, respectively. The changes of expression level of cell cycle genes in tumor cells were detected by reverse transcription polymerase chain reaction（RT- PCR）. Results： In situ hybridization analysis showed that the SPATA12 was highly expressed in normal adult testis, but lacking in testicular tumors such as seminoma. MTT assay and colony-forming assay indicated that the exogenous expression of SPATA12 could suppress both tumor cell proliferation and colony formation. RT-PCR showed that the expression of cyclin A1 gene was markedly suppressed and the level of cyclin D1 was somewhat reduced following SPATA12 transfection. However, no obvious changes were observed in mRNA expression of cyclin B1 or cyclin E1 after SPATA12 transfection. Conclusion： SPATA12 could be an inhibitor during the development of tumor via regulation of cell cycle genes.

关 键 词：精子发生相关基因12 肿瘤抑制 细胞周期基因 

分 类 号：R730.2[医药卫生—肿瘤]