摘要
目的:探讨肿瘤转移抑制基因KAI1基因对子宫内膜癌细胞(AN3CA和HEC-1-B)增殖及侵袭能力的影响。方法:脂质体介导pcDNA3-KAI1质粒转染人子宫内膜癌细胞AN3CA和HEC-1-B,采用免疫印迹法和流式细胞术检测转染前后肿瘤细胞KAI1蛋白的表达,MTT比色法、软琼脂克隆形成实验观察KAI1基因对肿瘤细胞增殖能力的影响,Transwell侵袭实验检测转染前后肿瘤细胞侵袭能力的变化。结果:转染pcDNA3-KAI1质粒后AN3CA和HEC-1-B细胞内和细胞表面均可检测到KAI1蛋白的稳定表达。转染空白质粒组细胞形成克隆数量多体积较大,细胞克隆形成率为(54.2±3.1)%和(52.7±4.3)%,细胞的倍增时间为21.3h和20.1h;基因转染pcDNA3-KAI1质粒组细胞形成克隆数少体积较小,细胞克隆形成率为(37.4±5.1)%和(32.1±3.7)%,细胞的倍增时间为43.7h和45.2h;两组间细胞增殖能力和克隆形成能力比较,差异均有统计学意义(P<0.05)。基因转染组细胞的穿膜细胞数为(91±10.7)/HT、(68±10.8)/HT,而空质粒转染组细胞的穿膜细胞数为(292±11.5)/HT、(219+12.7)/HT,两组细胞比较,侵袭能力亦显著下降(P<0.05)。结论:外源性肿瘤转移抑制基因KAI1有效转染可使子宫内膜癌细胞增殖、侵袭能力均下降,KAI1可能成为子宫内膜癌转移的新治疗靶点。
Objective： To investigate the influence of metastasis suppressor gene KAI1 on the proliferation, invasion and metastasis of endometrial carcinoma cell line AN3CA and HEC-1- B. Methods： The KAI1 cDNA was transfected into human endometrial carcinoma cells AN3CA and HEC-1-B via Lipofectamine 2000. The expression of KAI1 protein was examined by Western blotting and flow cytometry before and after transfection. The proliferation ability of AN3CA and HEC- 1-B cells was observed by MTT assay and anchorage-independent growth assay. The changes of cell invasive ability were studied by transwell assays. Results： Stable expression of KAI1 protein was observed in AN3CA and HEC-1- B cells and on their surface after transfection with pcDNA3-KAI1 plasmid. Cells transfected with blank plasmid formed more colonies and had a larger size, with the colony forming rates being（54.2 ± 3.1 ） % for AN3CA cells and （52.7±4.3 ） % for HEC- 1- B cells; the doubling time of AN3CA and HEC-1-B cells were 21.3 h and 20.1 h, respectively. Cells transfected with pcDNA3-KAI1 formed less colonies and had a smaller size, with the colony forming rates being （ 37.4 ±5. 1 ） % for AN3CA cells and （32.1±3.7）% for HEC-1- B cells; the doubling time of AN3CA and HEC-1-B cells were 43.7 h and 45.2 h, respectively. The cell proliferation abilities and colony-forming ability were significantly different between the two groups （P 〈 0.05 ）. The trans-membrane cell numbers in pcDNA3-KAI1 transfected AN3 CA and HEC-1-B cells were （91 ± 10.7）/HT and （68 ± 10.8 ）/HT, respectively; and in blank plasmid transfected cells were （292 ±11.5 ）/HT and（219 ± 12.7 ）/HT, respectively; there was significant difference between the two groups （P 〈0.05） . Conclusion： Exogenous metastasis suppressor KAI1 gene can effectively decrease the proliferation and invasive ability of human endometrial carcinoma cells ; KAI1 might be a potential target for treatment of metastasis of endometrial carcinoma.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2007年第5期445-449,共5页
Chinese Journal of Cancer Biotherapy
基金
国家重点基础研究发展(973)规划项目(No.2002CB513107)
国家自然科学基金项目(No.30528012)
海南省教育厅高校科研基金项目(No.HjKj200747)
Supported by the Major State Basic Research Development Program of China(No.2002CB513107)
National Natural Science Foundation of China(No.30528012)
the Research Fund of Higher Education of Hainan Ministry of Education(No.Hjkj200747)
