过表达ITLN-1能抑制人子宫内膜癌Ishikawa细胞的增殖并诱导其凋亡  

Over-expression of ITLN-1 inhibits proliferation and promotes apoptosis of human endometrial carcinoma Ishikawa cells

作　　者：姚兰[1] 陈书成[1] 

机构地区：[1]河北医科大学第四医院妇科,河北石家庄050011

出　　处：《肿瘤》2016年第5期496-503,共8页Tumor

基　　金：河北省医学科学研究重点课题计划资助项目(编号:20130233)~~

摘　　要：目的 :观察过表达肠凝集素1(intelectin 1,ITLN-1)对子人宫内膜腺癌Ishikawa细胞凋亡和增殖的影响。方法:采用脂质体法将重组质粒pCMV6-ITLN-1转入Ishikawa细胞(ITLN-1过表达组),同时设未做任何处理的细胞作为空白对照组和转入空载体pCMV6-N1的空载体组。48 h后,应用实时荧光定量PCR法检测ITLN-1、p53、环氧合酶2(cyclooxygenase-2,Cox-2)和第10号染色体缺失的磷酸酶及张力蛋白同源基因(phosphate and tension homology deleted on chromsome ten,PTEN)mRNA的表达水平;蛋白质印迹法检测ITLN-1、p53、Cox-2、Bcl-2及cleaved-caspase 3蛋白的表达水平。采用MTS法检测细胞的增殖活性;FCM法检测细胞的凋亡率,倒置相差显微镜下观察凋亡细胞形态的变化。结果:ITLN-1、p53和PTEN mRNA的表达水平均较空白对照组和空载体组明显上调(P<0.01、P<0.05和P<0.01),而Cox-2 mRNA的表达水平则明显下调(P<0.01)。p53和cleaved-caspase 3蛋白的表达水平均明显上调(P值均<0.01),Bcl-2和Cox-2蛋白的表达水平均明显下调(P值均<0.01)。与2个对照组相比,24和48 h时ITLN-1过表达组Ishikawa细胞的生长受到明显抑制(P<0.05和P<0.01),细胞凋亡率明显提高(P<0.01),倒置相差显微镜下细胞形态呈凋亡改变。结论 :ITLN-1表达上调能抑制Ishikawa细胞的增殖,并诱导其凋亡,这可能与上调Ishikawa细胞中凋亡相关蛋白的表达有关。Objective： To investigate the effects of over-expression of intelectin 1 （ITLN-1） on apoptosis and proliferation of human endometria carcinoma Ishikawa cells.Methods： The recombinant plasmid pCMV6-1TLN-1 was transfected into Ishikawa cells （named as ITLN-1 over-expression group）; at the same time, Ishikawa cells without transfection and transfected with empty vector pCMV6-N1 were used as the blank control group and empty vector group, respectively. The expression levels of ITLN-1, p53, cyclooxygenase-2 （Cox-2） and phosphate and tension homology deleted on chromosome ten （PTEN） mRNAs were detected by real-time fluorescent quantitative PCR. The expression levels of ITLN-1, p53, Cox-2, Bcl-2 and cleaved-caspase 3 proteins were detected by Western blotting. The cell proliferation was measured by MTS method. The apoptosis rate was measured by FCM. The morphological changes of apoptotic cells were observed under an inverted phase contrast microscope. Results： The expression levels of ITLN-1, p53 and PTEN mRNAs in Ishikawa cells transfected with ITLN-1 over-expression plasmid were significantly up-regulated as compared with those in the blank control group and empty vector group （P 〈 0.01, P 〈 0.05 and P 〈 0.01）, but the expression level of Cox-2 mRNA was significantly down-regulated （P 〈 0.01）. The expression levels of p53 and cleaved-caspase 3 proteins were significantly up-regulated （both P 〈 0.01）, while the expression levels of Cox-2 and Bcl-2 proteisn were significantly down-regulated （both P 〈 0.01）in ITLN-1 over-expression group. Compared with the two control groups, the cell proliferation of Ishikawa cells transfected with ITLN-1 over-expression plasmid at 24 and 48 h was significantly inhibited （P 〈 0.05, P 〈 0.01）, the apoptosis rate was obviously increased （P 〈 0.01）, meanwhile the cellular morphology showed the characteristics of apoptosis under the inverted phase contrast microscope. Conclusion： Up-regulation of ITLN-1 expression can inhibit

关 键 词：子宫内膜肿瘤 凝集素类 细胞增殖 细胞凋亡 

分 类 号：R737.33[医药卫生—肿瘤]