肿瘤转移抑制基因KAI1对子宫内膜癌细胞增殖、侵袭能力的影响

Influence of metastasis suppressor gene KAI1 on proliferation and invasion of endometrial carcinoma cells

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.