肾癌荷瘤小鼠组织中补体 C5a / C5aR 通路活化与肿瘤高凝状态的关系及其机制研究

目的 研究肾癌荷瘤小鼠组织中补体 C5a / C5aR 通路活化与肿瘤高凝状态的关系及其机制。 方法 收集 2016 年 7 月至 2019 年 10 月于唐山市人民医院收治的 32 例经病理学活检确诊的肾癌患者的癌组织和癌旁组织。 采用 Western 印迹法检测组织中 C5a / C5aR 通路活化相关蛋白表达情况。 于 BALB/ c-nu / nu 裸鼠前肢腋窝皮下接种人 ACHN 肾癌细胞株, 接种成功后随机分为干预组、 模型组, 其中干预组尾静脉注射C5a 联合 C5aR 阻断剂 (C5aR antagonist, C5aRA), 模型组等体积注射生理盐水, 另取 6 只空白小鼠作为对照组。 观察小鼠肿瘤生长情况, 检测高凝状态相关指标水平。 脱颈处死小鼠, 收集癌组织, 采用 Western印迹法检测组织中 C5a / C5aR 通路活化相关蛋白表达情况。 结果 两组小鼠肿瘤体积于荷瘤 6 ~ 21 d 差异具有统计学意义, 且干预组的肿瘤体积明显低于模型组 ( P< 0. 05)。 3 组小鼠于荷瘤第 1 天的血清 D-D、vWF、 TF 水平差异无统计学意义, 荷瘤第 14 天、 第 21 天干预组、 模型组血清凝血指标水平明显增加, 其中干预组均明显低于模型组 (P< 0. 05)。 干预组癌组织中 C5aR、 C5b-9、 FGL2、 P38、 p-P38 的表达水平均明显低于模型组 (P< 0. 05)。 肾癌患者癌组织中 C5aR、 C5b-9、 FGL2、 P38、 p-P38 的表达水平均明显高于癌旁组织 (P< 0. 05)。 32 例肾癌患者中高凝状态患者 14 例, 非高凝状态患者 18 例, 高凝状态组癌组织中C5aR、 C5b-9、 FGL2、 P38、 p-P38 的表达水平均明显高于非高凝状态组 (P< 0. 05)。 结论 补体 C5a / C5aR通路可通过调控 FGL2 的表达而诱导肾癌的高凝状态, 进而参与肾癌的发病过程。

Relationship between Activation of Complement C5 a/C5 aR Pathway and Tumor Hypercoagulability in Renal Carcinoma-bearing Mice

Objective To investigate the relationship between the activation of complementC5a / C5aR pathway and tumor hypercoagulability and the underlying mechanism in renal carcinoma bearing mice. Methods The cancer and paracancerous tissues of 32 patients with renal cell carcinoma diagnosed by pathological biopsy in the Tangshan People’s Hospital from July 2016 to October2019 were obtained. The expression levels of C5a / C5aR pathway activation related proteins in tissueswere detected by Western blotting. The human ACHN renal cancer cells were inoculated subcutaneously in the axillary region of the BALB / c-nu / nu nude mice. After successful inoculation, they wererandomly divided into intervention group and model group, in which the intervention group was injected with C5aR antagonist (C5aRA) via the tail vein, and the model group was injected withnormal saline in equal volume, and 6 mice with no subcutaneous inoculation and no C5aRA treatment were taken as the control group. The tumor growth was observed and the levels of hypercoagulability related indexes were measured. The mice were sacrificed to collect cancer tissues, and the expression levels of C5a / C5aR pathway activation related proteins were detected by Western blotting. Results The tumor volume of mice in the two groups was statistically significant in 6 ~ 21 day,and the tumor volume of the intervention group was significantly lower than that of the model group(P< 0. 05). There was no significant difference in serum D-D, vWF and TF levels in the first day oftumor-bearing mice in the three groups. The serum coagulation index levels in the intervention groupand the model group were significantly increased at the 14th day and the 21st day. Moreover, the serum coagulation index levels in the intervention group were significantly lower than those in the model group (P< 0. 05). The expression levels of C5aR, C5b-9, FGL2, P38 and p-P38 in mice cancer tissues in the intervention group were significantly lower than those in the model group ( P <0. 05). The expression levels of C5aR, C5 b-9, FGL2, P38 and p-P38 in renal carcinoma weresignificantly higher than those in adjacent tissues (P< 0. 05). There were 14 cases of hypercoagulability and 18 cases of non-hypercoagulability in 32 cases of renal carcinoma. The expression levels ofC5aR, C5 b-9, FGL2, P38 and p-P38 in hypercoagulable group were significantly higher thanthose in non-hypercoagulable group (P< 0. 05). Conclusion Complement C5a / C5aR pathway caninduce hypercoagulability of renal carcinoma by regulating the expression of FGL2, and then participate in the pathogenesis of renal carcinoma.