重组腺病毒Ad-IL-12感染淋巴细胞对人卵巢癌细胞SKOV3增殖和凋亡的影响  

Effects on proliferation and apoptosis of ovary cancer cells SKOV3 co-cultrued with lymphocytes infected by recombinant adenovirus Ad-IL-12

作　　者：王秦[1] 成凤[1] 匡文斌[1] 董晋豫[1] 马婷婷[1] 熊海玉[1] 涂植光[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点试验室,重庆400016

出　　处：《基础医学与临床》2013年第8期947-954,共8页Basic and Clinical Medicine

基　　金：国家自然科学基金(81172016)

摘　　要：目的探讨重组腺病毒Ad-IL-12感染人外周血淋巴细胞后对人卵巢癌细胞SKOV3周期、增殖和凋亡的影响。方法采用重组人IL-12腺病毒(Ad-IL-12)感染人外周血淋巴细胞,感染48 h后与人卵巢癌细胞SKOV3共培养(SKOV3/Ad-IL-12),同时设未感染组(SKOV3)和Ad-GFP空载感染组(SKOV3/Ad-GFP)作为对照。RT-PCR检测IL-12双亚基P35和P40的mRNA表达,ELISA检测细胞培养上清中IL-12 P70蛋白的分泌,CCK8法检测卵巢癌细胞增殖,流式细胞术分析卵巢癌细胞周期和凋亡,RT-PCR和Western blot检测抗凋亡蛋白BCL-2、survivin的表达。结果重组腺病毒Ad-IL-12可成功感染人外周血淋巴细胞,分泌较高水平的IL-12 P70蛋白;淋巴细胞过表达IL-12可抑制SKOV3细胞的增殖;与未感染组和空载组相比,SKOV3/Ad-IL-12组G1期细胞显著增加,凋亡率显著升高(P<0.05),SKOV3细胞中抗凋亡蛋白BCL-2和survivin显著下调(P<0.05)。结论人IL-12基因重组腺病毒可以成功感染人外周血淋巴细胞,分泌IL-12 P70蛋白,并可以通过阻滞细胞周期进程和促进细胞凋亡抑制人卵巢癌细胞SKOV3的增殖。Objective To investigate the effects of lymphocytes infected by recombinant adenovirus Ad-IL-12 on the proliferation and apoptosis of ovary cancer cell SKOV3. Methods The peripheral blood lymphocytes were infected by recombinant adenovirus Ad-IL-12 and Ad-GFP for 48 h. The SKOV3 cells were co-cultured with lymphocytes in- fected with Ad-IL-12 （SKOV3/Ad-IL-12） and lymphocytes infected with Ad-GFP （SKOV3/Ad-GFP） separately, and the untreated cells （SKOV3） were set as another control. IL-12 double-subunits （P35, P40） mRNA expres- sions were tested by RT-PCR and the level of IL-12PTO protein in the supernatant was detected with ELISA. The proliferation potential of SKOV3 was assessed by CCK8 assay, flow cytometry was used to detect the cell cycle and cellular apoptosis, RT-PCR and Western blot were used to detect the expressions of BCL-2 and survivin. Results The human recombinant adenovirus Ad-IL-12 successfully infected the human peripheral blood lymphocytes and ef- fectively secreted IL-12P70 proteins; growth ability of SKOV3 cells （SKOV3/Ad-IL-12） was significantly inhibited （P 〈0. 05 ）. In addition, the percentages of SKOV3 cells in the G1 phase and the apoptosis （SKOV3/Ad-IL-12） increased Significantly （ P 〈 0. 05 ）, the expression of BCL-2 and survivin were down-regulated （ P 〈 0. 05 ）. Con- clusions The human recombinant adenovirus Ad-IL-12 could infect the human peripheral blood lymphocytes and express the IL-12 P70 protein successfully, which may inhibit the proliferation of SKOV3 cells by blocking cell cy- cle and inducing apoptosis.

关 键 词：Ad—IL-12 人卵巢癌细胞SKOV3 增殖 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]