[卵巢癌细胞抑制T细胞JAK-STAT信号转导通路的实验研究]

[Ovarian carcinoma cell inhibits T cell JAK-STAT signal transduction pathway, an experimental study].

作者信息

Wang Hui, Xie Xing, Lü Wei-guo, Ye Da-feng, Chen Huai-zeng, Li Xiao, Cheng Qi

机构信息

Women's Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2003 Jun 10;83(11):972-5.

PMID:12899799

Abstract

OBJECTIVE

To investigate the effect of ovarian carcinoma cell on T cell JAK-STAT signal transduction pathway and its role in the ovarian carcinoma induced immunosupression.

METHODS

Human ovarian carcinoma cells of OVCAR3. CAOV3, and SKOV3 lines were cultured. CD8(+) T cells were isolated from the peripheral venous blood of healthy persons. Then the supernatants of these ovarian carcinoma cell lines and RPMI-1640 were added into the culture of CD8(+) T cells (groups I, II, III, and control). Thiazolyl blue (MTT) method was used to detect the growth of CD8(+) T cell. The cell cycle was examined by flow cytometry. The secretion of the Tc1 type cytokine interferon (IFN)-gamma mRNA and the secretion of the Tc2 type cytokine interleukin (IL)-10 mRNA were detected by RT-PCR. The expression of signaling molecules JAK and JAK3 and the phosphorylated activation of STAT3 and STAT5 in the CD8(+) T cell were analyzed by Western blotting.

RESULTS

The absorbance at the wavelength 570 nm of CD8(+) T cell culture was 0.23 +/- 0.03, 0.28 +/- 0.06, and 0.29 +/- 0.05 in the group I, II, and III, all significantly lower than that in the group IV (0.79 +/- 0.07, all P < 0.01). The percentages of CD8(+) T cells at the stage S and stage G(2)/M were lower, and those in stage G(1)/G(0) were higher in groups I, II, and III than in group IV (all P < 0.01). The IFN-gamma expression was significantly lower in groups I, II, and III in comparison with that in group IV. However, the expression of IL-10 was significantly higher in groups I, II, and III in comparison with that in group IV. The expression of JAK3 protein, but not JAK1 protein, was significantly lower in groups I, II, and III in comparison with that in group IV. The phosphorylated activation of STAT5 was suppressed significantly in groups I, II, and III, whereas the phosphorylated activation of STAT3 was suppressed only in group I.

CONCLUSION

Ovarian carcinoma may suppress T cell proliferation through inhibition of the JAK-STAT signal transduction pathway, which may be a mechanism of ovarian carcinoma induced immunosuppression.

摘要

目的

探讨卵巢癌细胞对T细胞JAK-STAT信号转导通路的影响及其在卵巢癌诱导免疫抑制中的作用。

方法

培养人卵巢癌细胞系OVCAR3、CAOV3和SKOV3。从健康人外周静脉血中分离CD8(+)T细胞。然后将这些卵巢癌细胞系的上清液和RPMI-1640加入CD8(+)T细胞培养物中（第I、II、III组和对照组）。采用噻唑蓝（MTT）法检测CD8(+)T细胞的生长情况。通过流式细胞术检测细胞周期。采用逆转录-聚合酶链反应（RT-PCR）检测Tc1型细胞因子干扰素（IFN）-γmRNA的分泌及Tc2型细胞因子白细胞介素（IL）-10mRNA的分泌。通过蛋白质印迹法分析CD8(+)T细胞中信号分子JAK和JAK3的表达以及STAT3和STAT5的磷酸化激活情况。

结果

第I、II、III组CD8(+)T细胞培养物在波长570nm处的吸光度分别为0.23±0.03、0.28±0.06和0.29±固定，均显著低于第IV组（0.79±0.07，均P<0.01）。第I、II、III组处于S期和G(2)/M期的CD8(+)T细胞百分比低于第IV组，而处于G(1)/G(0)期的高于第IV组（均P<0.01）。与第IV组相比，第I、II、III组中IFN-γ的表达显著降低。然而，与第IV组相比，第I、II、III组中IL-10的表达显著升高。与第IV组相比，第I、II、III组中JAK3蛋白的表达显著降低，而JAK1蛋白的表达无明显变化。第I、II、III组中STAT5的磷酸化激活明显受到抑制，而仅第I组中STAT3的磷酸化激活受到抑制。