基础医学与临床 ›› 2013, Vol. 33 ›› Issue (8): 947-954.

• 研究论文 • 上一篇    下一篇

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

王秦,成凤,匡文斌,董晋豫,马婷婷,熊海玉,涂植光   

  1. 重庆医科大学检验医学院
  • 收稿日期:2012-08-01 修回日期:2012-10-25 出版日期:2013-08-05 发布日期:2013-07-18
  • 通讯作者: 涂植光 E-mail:cqwq1986925@sina.com
  • 基金资助:
    靶向肿瘤相关巨噬细胞过表达IL-12诱导其表型和功能改变及其分子机制探讨

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

  • Received:2012-08-01 Revised:2012-10-25 Online:2013-08-05 Published:2013-07-18
  • Contact: Zhi-Guang TU E-mail:cqwq1986925@sina.com

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增殖、周期和凋亡的影响。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。

关键词: Ad-IL-12, 人卵巢癌细胞SKOV3, 增殖, 凋亡

Abstract: 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 respectively.The SKOV3 cells was co-cultured with lymphocytes infected 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 expressions were tested by RT-PCR and the levels of IL-12P70 protein in the cell culture supernatant were 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 expression of BCL-2 and Survivin. Results The human recombinant adenovirus Ad-IL-12 could successfully infect the human peripheral blood lymphocytes and effectively secret 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) were increased Significantly(P<0.05), the expression of BCL-2 and Survivin were down-regulated(P<0.05). Conclusions The human recombinant adenovirus Ad-IL-12 could infect the human peripheral blood lymphocytes and express the IL-12 P70 protein successfully, which could inhibit the proliferation of SKOV3 cells by blocking cell cycle and inducing apoptosis.

Key words: Ad-IL-12, human ovary cancer cells SKOV3, proliferation, apoptosis

中图分类号: