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目的 探讨两株人胆管癌细胞系中自分泌性白细胞介素 6(IL 6)的作用以及生长抑素类似物奥曲肽对其的影响。方法 以酶联免疫吸附试验测定胆管癌细胞系IL 6的分泌 ,IL 6受体亚单位 gp80和 gp1 30mRNA的表达及表达水平变化以逆转录 聚合酶链式反应 (RT PCR)检测。体外IL 6对胆管癌细胞增殖的作用应用噻唑蓝 (MTT)比色法检测。结果 无血清培养条件下胆管癌细胞株NEC和SSP 2 5可以分泌IL 6 ,分别为 (2 67.0± 2 6 .3)ng/L和 (775 .5± 91 .0 )ng/L ,奥曲肽 (OCT ,1mg/L)作用 2 4h可以抑制这两种胆管癌IL 6的分泌 ,分别下降至 (1 86 .5± 2 9.7)ng/L和 (31 8.6± 1 1 5 .0 )ng/L(P <0 .0 1 )。两种胆管癌细胞株皆可表达IL 6的受体亚单位 gp80和gp1 30mRNA ,但计算积分吸光度比值显示奥曲肽对IL 6受体mRNA的表达无影响。IL 6(1 0 μg/L)可以促进胆管癌细胞系NEC和SSP 2 5的增殖 (A值分别为 0 .488± 0 .0 4 8、0 .699± 0 .0 30 ,对照组为 0 .383± 0 .0 2 7、0 .41 3± 0 .0 52 ,P <0 .0 1 ) ,奥曲肽则可以明显抑制IL 6刺激的增殖 (0 .376±0 .0 2 3、0 .441± 0 .0 2 4 ,与IL 6组比较 ,P <0 .0 1 )。结论 胆管癌细胞系NEC和SSP 2 5中存在IL 6自分泌促生长环路 ,奥曲肽可以减少这两种胆管癌细胞分泌IL 6 ,?
Objective To investigate the role of autocrine interleukin 6 (IL 6) in two human cholangiocarcinoma cell lines and the effect of somatostatin analogue octreotide on it. Methods The secretion of IL 6 in cholangiocarcinoma cell lines was determined by enzyme-linked immunosorbent assay. The expression levels of gp80 and gp1 30 mRNA in IL 6 receptor subunits were detected by reverse transcription polymerase chain reaction (RT PCR). The effect of IL 6 on proliferation of cholangiocarcinoma cells in vitro was detected by MTT assay. RESULTS: Serum-free cultured cholangiocarcinoma cell lines NEC and SSP 25 secreted IL 6, which were (2 67.0 ± 2 6.3) ng/L and (775. 5 ± 91.0) ng/L, respectively. OCT, 1mg/L) for 24h inhibited the secretion of IL 6 in these two types of cholangiocarcinoma and decreased to (1 86. 5± 2 9.7) ng/L and (31 8.6± 1 1.5) ng/L, respectively. (P <0. 0 1). Both bile duct cancer cell lines can express IL 6 receptor subunits gp80 and gp1 30 mRNA, but calculating the integral absorbance ratio shows that octreotide has no effect on IL 6 receptor mRNA expression. IL 6 (10 μg/L) promoted the proliferation of cholangiocarcinoma cell lines NEC and SSP 25 (A values were 0.488 ± 0.048, 0.69 ± 0.003, respectively, and 0 in the control group. The results showed that octreotide could significantly inhibit the proliferation of IL-6 stimulation (0.376±0. 0 2 3, 0. 383 ± 0. 0 27, 0. 41 3 ± 0. 0 52, P <0. 01). 441 ± 0 .0 2 4 compared with IL 6 group, P <0 .0 1 ). Conclusion There is an IL-6 autocrine growth-promoting loop in the bile duct cancer cell lines NEC and SSP 25, and octreotide can reduce the secretion of IL 6 from these two types of cholangiocarcinoma cells.