卵巢上皮性恶性肿瘤患者T淋巴细胞相关细胞因子与顺铂耐药的相关性

Relationship between T lymphocyte-associated cytokines and cisplatin resistance in patients with ovarian epithelial carcinoma

目的探讨卵巢上皮性恶性肿瘤患者血清、外周血单个核细胞(PBMC)和肿瘤组织中T淋巴细胞相关细胞因子IL-2、IL-4、IL-10、TNF-α和IFN-γ表达与顺铂耐药的相关性。方法采用ATP生物荧光肿瘤体外药物敏感检测(ATP-TCA)法检测37例卵巢上皮性恶性肿瘤患者肿瘤组织的顺铂耐药性,根据患者对顺铂的敏感和耐药情况将其分为耐药组(11例)和敏感组(26例)。采用ELISA法分别检测两组患者血清中IL-2、TNF-α和IFN-γ水平,采用实时定量PCR检测两组患者PBMC和肿瘤组织中IL-2、IL-4、IL-10、TNF-α和IFN-γ的mRNA表达水平,应用流式细胞仪检测PBMC和CD4^+、CD8^+T淋巴细胞中IFN-γ蛋白质表达水平。结果耐药组血清IL-2和IFN-γ水平均显著低于敏感组(P值均<0.05),两组间血清TNF-α水平的差异无统计学意义(P>0.05)。敏感组肿瘤组织中TNF-α和IFN-γ的mRNA表达水平均显著高于耐药组(P值均<0.05),两组间IL-2、IL-4、IL-10的mRNA表达水平的差异均无统计学意义(P值均>0.05);敏感组PBMC中IFN-γ的mRNA表达水平显著高于耐药组(P<0.05),IL-2、IL-4、IL-10和TNF-α的mRNA表达水平较耐药组有升高的趋势,但差异均无统计学意义(P值均>0.05)。两组PBMC中IFN-γ蛋白质表达水平的差异无统计学意义(P>0.05),敏感组CD4^+T和CD8^+T淋巴细胞中的IFN-γ蛋白质表达水平均显著高于耐药组(P值分别<0.01、0.05)。结论顺铂耐药性与CD4^+、CD8^+T淋巴细胞的免疫应答状态相关,IFN-γ可能在这个过程中发挥重要作用。

Objective To examine the expression of T lymphocyte-associated cytokines including interleukin(IL)-2, IL-4, IL-10, interferon(IFN)-α, and IFN-γ in the serum, peripheral blood mononuclear cells(PBMC) and tumor tissues from cisplatin-resistant ovarian cancer patients, and to analyze the role of the cytokines in the cisplatin resistance. Methods Adenosine triphosphate-tumor chemosensitivity assay(ATP-TCA) was used to detect the cisplatin resistance in 37 ovarian cancer patients. Of the 37 patients, 11 were resistant to cisplatin(cisplatin-resistant group) and 26 were sensitive to cisplatin(sensitive group). Enzyme-linked immunosorbent assay(ELISA) was used to detect serum IL-2, tumor necrosis factor(TNF)-α and IFN-γ. Real time polymerase chain reaction(RT-PCR) was used to determine mRNA levels of IL-2, IL-4, IL-10, TNF-α and IFN-γ in PBMC and tumor tissues. Flow cytometry was used to test IFN-γ protein level in PBMC, CD4^+ and CD8^+T cells. Results Serum levels of IL-2 and IFN-γ in cisplatin-resistant group were significantly lower than those in sensitive group(both P<0.05), while there was no significant difference in the TNF-α expression between two groups(P>0.05). The mRNA levels of TNF-α and IFN-γ in tumor tissues in sensitive group were significantly higher than those in cisplatin-resistant group(P<0.05), there were no significant differences in the IL-2, IL-4 or IL-10 mRNA levels between two groups(all P>0.05). IFN-γ mRNA level was significantly upregulated in PBMC from cisplatin-sensitive group as compared with cisplatin-resistant group(P<0.05). There was a slight trend of upregulations of IL-2, IL-4, IL-10 and TNF-α mRNA levels in PBMC from cisplatin-sensitive group, but no significant difference was found between groups(all P>0.05). There was no significant difference in IFN-γ protein expression in PBMC between two groups(P>0.05), while IFN-γ protein expressions in CD4^+ and CD8^+T cells in sensitive group were significantly higher than those in cisplatin-resistant group(P<0.01, 0.05). Conclusion Cisplatin sensibility is correlated with immune response condition of CD4^+ and CD8^+T cells. IFN-γ may participate in the cisplatin-resistant process.