过表达JAZF1对甲状腺乳头状癌BCPAP细胞增殖、凋亡的影响及其机制

Effect of JAZF1 overexpression on proliferation and apoptosis in papillary thyroid carcinoma BCPAP cells and its mechanism

目的探讨相互并列的锌指基因1(juxtaposed with another zinc finger gene 1,JAZF1)对甲状腺乳头状癌(papillary thyroid cancer,PTC)BCPAP细胞增殖、凋亡的影响及可能的作用机制。方法采用重组腺病毒Adv-JAZF1-GFP、Adv-GFP分别感染BCPAP细胞,分为阴性对照组(NC组)、空载组(Adv-GFP组)、实验组(Adv-JAZF1-GFP组),感染48 h后分别提取各组细胞总RNA、蛋白,采用实时荧光定量PCR(qRT-PCR)、Western blot检测JAZF1 mRNA、蛋白表达水平,MTT、克隆形成实验检测细胞生长及增殖能力,流式细胞术annexin V-FITC/PI双染检测各组细胞凋亡情况,Western blot检测分析Bcl-2、Bax、PPAR-γ、PI3K、Akt、p-Akt蛋白表达水平。结果与阴性对照组、空载组比较,实验组JAZF1 mRNA、蛋白表达水平均明显上调(P<0.001),且随时间增殖能力明显抑制(P<0.01),克隆形成能力降低(P<0.001);流式annexin V-FITC/PI双染检测结果显示实验组细胞凋亡率[(23.02±0.35)%]较NC组[(8.63±0.40)%]、空载组[(7.95±0.32)%]均明显增加(P<0.001);Western blot结果显示实验组Bcl-2表达较NC组、空载组明显下调66.0%(P<0.001),Bax表达明显上调64.8%(P<0.001),并显著降低p-PI3K/PI3K、pAkt/Akt比值(P<0.001)。结论JAZF1通过调控Bcl-2/Bax通路促进细胞凋亡,并且调控PI3K/Akt信号通路抑制BCPAP细胞增殖,从而在甲状腺乳头状癌的发展进程中可能发挥重要作用。

Objective To investigate the effect of juxtaposed with another zinc finger 1(JAZF1)on proliferation and apoptosis of papillary thyroid carcinoma(PTC)BCPAP cells and its possible mechanism.Methods BCPAP cells were divided into negative control group(NC group),Adv-GFP group and Adv-JAZF1-GFP group,which were not infected or infected with recombinant adenoviruses Adv-GFP or Adv-JAZF1-GFP,respectively.The total RNA and protein of each group were extracted 48 h after infection.The relative JAZF1 mRNA and protein expressions were determined by real-time fluorescence quantitative PCR(qRT-PCR)and Western blotting,and the growth and proliferation of cells were examined by the MTT and clone formation assays.Annexin V-FITC/PI double staining was used to determine the apoptosis of cells in each group by flow cytometry.The relative protein expressions of Bcl-2,Bax,PPAR-γ,PI3K,Akt and p-Akt were determined by Western blotting.Results Compared with the NC and Adv-GFP groups,the relative JAZF1 mRNA and protein expression in the experimental group was significantly upregulated(P<0.001),the proliferation ability was significantly inhibited(P<0.01),and the clone formation ability was decreased(P<0.001).The Annexin V-FITC/PI double staining results showed that the apoptosis rate of the Adv-JAZF1-GFP group(23.02±0.35%)was significantly increased than the NC group(8.63±0.40%)and the Adv-GFP group(7.95±0.32%)(P<0.001).Compared with NC and Adv-GFP groups,the expression level of Bcl-2 was significantly downregulated by 66.0%and the expression of Bax was upregulated by 64.8%in the Adv-JAZF1-GFP group(P<0.001).Furthermore,the p-PI3K/PI3K and pAkt/Akt ratios were significantly decreased in the experimental group compared with the controls(P<0.001).Conclusions JAZF1 may play an important role in the development of papillary thyroid cancer by regulating the Bcl-2/Bax pathway to promote apoptosis and by regulating the PI3K/Akt signaling pathway to inhibit the proliferation of BCPAP cells.