直肠癌组织中嘌呤能离子通道型受体7表达的意义及其与肿瘤细胞特性的关系

Significance of P2X7R Expression in Rectal Cancer and Relationship with Tumor Cell Characteristics

目的:探讨直肠癌组织中嘌呤能离子通道型受体7(P2X7R)表达与肿瘤病理分期、脉管浸润及肿瘤细胞增殖、侵袭能力的关系。方法:收集2019年10月—2020年10月在我院就诊的直肠癌患者的癌组织标本120例,同时选取癌旁组织作为对照,采用免疫组化染色法检测P2X7R表达;购买人结直肠癌细胞株HCT116及正常结直肠上皮细胞NCM460,将HCT116细胞随机分为对照组、P2X7R激动剂低剂量组(0.01 mmol/L)、P2X7R激动剂高剂量组(0.1 mmol/L),其中P2X7R激动剂组给予苯甲酰-4-苯甲酰-三硝基苯磺酸(BzATP),采用CCK法检测细胞增殖情况,侵袭实验检测细胞侵袭能力,Western blotting检查p-STAT3、PWX7R蛋白相对表达量。结果:直肠癌组织P2X7R阳性表达率为37.50%,明显低于癌旁组织(91.67%),差异有统计学意义(P<0.05);TNM分期Ⅲ~Ⅳ期、有脉管浸润直肠癌组织P2X7R阳性表达率为18.60%和16.67%,明显低于Ⅰ~Ⅱ期、无脉管浸润直肠癌组织(P<0.05);HCT116细胞PWX7R蛋白相对表达量为(0.233±0.082),明显低于NCM460(0.982±0.102)细胞,差异有统计学意义(P<0.05);0.01 mmol/L激动剂组、0.1 mmol/L激动剂组细胞培养12 h、24 h和48 h时OD值明显低于对照组,0.1 mmol/L激动剂组细胞培养12 h、24 h和48 h时OD值分别为(0.300±0.096)、(0.446±0.106)和(0.570±0.110),明显低于0.01 mmol/L激动剂组,差异有统计学意义(P<0.05);0.01 mmol/L激动剂组、0.1 mmol/L激动剂组细胞侵袭数低于对照组,0.1 mmol/L激动剂组细胞侵袭数为(68.38±20.11)个,明显低于0.01 mmol/L激动剂组,差异有统计学意义(P<0.05);0.01 mmol/L激动剂组、0.1 mmol/L激动剂组p-STAT3、PWX7R蛋白相对表达量高于对照组,0.1 mmol/L激动剂组p-STAT3、PWX7R蛋白相对表达量分别为(1.322±0.143)和(1.011±0.132),明显低于0.01 mmol/L激动剂组,差异有统计学意义(P<0.05)。结论:直肠癌P2X7R表达下调,与肿瘤分期及脉管浸润有关,促进P2X7R表达可抑制直肠癌细胞增殖及侵袭能力。

Objective To investigate the expression of Purinergic P2X7 receptor(P2X7R)in rectal cancer and its relationship with tumorigenesis,cell proliferation and invasion.Methods 120 patients with rectal cancer in our hospital from October 2019 to October 2020 were selected and the adjacent tissues were selected as control.The expression of P2X7R was detected by immunohistochemistry.The human colorectal cancer cell line HCT116 and normal colorectal epithelial cell line NCM460 were selected,and HCT116 cells were randomly divided into control group,which 0.01 mmol/L agonist group and 0.1 mmol/L agonist group.The agonist group was given benzoyl-4-benzoyl-trinitrobenzene sulfonic acid,and MTT assay was used to detect cell proliferation.The invasion ability of the cells was detected by invasion assay.The Western blotting was used to detect the relative expression of p-STAT3 and PWX7R.Results The positive expression rate of P2X7R in rectal cancer was 37.50%,which was significantly lower than that in adjacent tissues(P<0.05).The positive expression rate of P2X7R in TNM stageⅢ-Ⅳand vascular invasion of rectal cancer was 18.60%and 16.67%,which were significantly lower than those in rectal cancer tissues with stageⅠ-Ⅱand without vascular invasion(P<0.05).The relative expression of P2X7R protein in HCT116 cells was(0.233±0.082),which was significantly lower than that in NCM460 cells(P<0.05).The OD values of 0.01 mmol/L agonist group and 0.1 mmol/L agonist group at 12 h,24 h and 48 h after cultured were significantly lower than those of control group(P<0.05).The OD values of 0.1 mmol/L agonist group at 12 h,24 h and 48 h after cultured were(0.300±0.096),(0.446±0.106)and(0.570±0.110)respectively,which were significantly lower than those of 0.01 mmol/L agonist group(P<0.05).The number of invasive cells in 0.01 mmol/L agonist group and 0.1 mmol/L agonist group were significantly lower than those in control group(P<0.05).The number of invasive cells in 0.1 mmol/L agonist group was(68.38±20.11),which was significantly lower than that in 0.01 mmol/L agonist group(P<0.05).The relative expression levels of p-STAT3 and P2X7R in 0.01 mmol/L agonist group and 0.1 mmol/L agonist group were higher than those in control group(P<0.05).The relative expression levels of p-STAT3 and P2X7R in 0.1 mmol/L agonist group were(1.322±0.143)and(1.011±0.132)respectively,which were significantly lower than those in 0.01 mmol/L agonist group(P<0.05).Conclusion The down-regulation of P2X7R expression in rectal cancer is related to tumor stage and vascular invasion,and P2X7R expression can inhibit the proliferation and invasion of rectal cancer cells.