p15^(INK4B)基因转染联合TGF-β_1对人食管鳞癌的抑制作用  

The Inhibitory Action of p15^(INK4B) Gene Transfection plus TGF-β_1 on Human Esophageal Squamous Cell Carcinoma Cell Line EC109

作　　者：张学彦[1] 刘铁夫[2] 于旸[3] 崔希威[1] 

机构地区：[1]哈尔滨医科大学附属第二医院消化科,哈尔滨市150086 [2]哈尔滨医科大学附属第四医院消化科 [3]哈尔滨医科大学遗传学教研室

出　　处：《中国肿瘤临床》2006年第20期1165-1168,共4页Chinese Journal of Clinical Oncology

基　　金：黑龙江省科技计划攻关重点项目基金(编号:GB05C401-09);黑龙江省卫生厅科研基金资助(编号:2004-117)

摘　　要：目的:探讨p15INK4B基因转染与TGF-β1联合应用对人食管鳞癌细胞系EC109细胞增殖和凋亡的影响。方法:脂质体介导将pCDNA3.1(+)-p15转染EC109细胞,稳定筛选后加入10ng/ml的TGF-β1。PCR检测外源p15基因,Westernblot检测转染细胞P15蛋白的变化。用MTT、集落形成实验、流式细胞仪和透射电镜检测p15转染TGF-β1对EC109增殖和凋亡的影响。结果:联合应用与二者单独应用相比,明显抑制EC109生长速度,集落形成率显著降低(P<0.01),发生G1/S阻滞,G1期比例显著升高(P<0.05),S期比例明显降低(P<0.05),凋亡率明显升高(P<0.01)。透射电镜发现二者联合应用诱导EC109发生明显的细胞凋亡。结论:p15基因转染与TGF-β1联合应用可以进一步增强对食管鳞癌EC109细胞的抑制作用并诱导其凋亡。Objective： To investigate the effect of combining p15 gene transfection with TGF-β1 on the proliferation and apoptosis of human esophageal squamous cell carcinoma cell line ECI09. Methods： The plasmid pCDNA3.1 （＋）-P15 was transfectcted into EC109 cell by the vector of liposomes and 10ng/mL TGF-β1 was added after stable sieving to examine the exogenous p15 gene. The western blot method was used to determine the change of p15 proteins. MTT, colony formation, flow cytometry and transmission electron microscope （TEM） were used for determination of the effect of p15 transfecting TGF-β1, on the proliferation and apoptosis of EC109. Results： Compared with the single application, the combination obviously inhibited the growth velocity and the colony formation rate was apparently decreased （P〈0.01）; limpid G1/S occurred clearly, the proportion of G1 phase was significantly increased （P〈0.05）, that of S phase was significantly decreased （P〈0.05）, and the rate od apoptosis was significantly increased （P〈0.01） compared with either p15 transfected group or added TGF-β1, group. It was also found under the transmission electron microscope that obvious apoptosis occurred in the combined p15 gene transfection with TGF-β1 group. Conclusion： p15 Combination of p15 gene transfection with TGF-β1, can further strengthen the inhibition and can induce the apoptosis on EC109 cells in vitro.

关 键 词：P15^INK4B 基因转染 TGF-β1食管鳞癌 

分 类 号：R735.1[医药卫生—肿瘤]