胃贲门腺癌组织PTPN12表达生物学意义研究  

Expression of PTPN12in gastric cardia adenocarcinoma and its effect on biological characteristics of gastric cancer cells

作　　者：牛云峰 王明辉 沈素朋[1] 梁佳[1] 郭炜[1] 董稚明[1] NIU Yun-feng;WANG Ming-hui;SHEN Su-peng;LIANG Jia;GUO Wei;DONG Zhi-ming(Laboratory of Pathology,Hebei Cancer Institute,Fourth Hospital of Hebei Medical University,Shijiazhuang050011,P.R.China;School of Nursing,Hebei College of Traditional Chinese Medicine,Shijiazhuang050200,P.R.China)

机构地区：[1]河北医科大学第四医院,河北省肿瘤研究所病理研究室,河北石家庄050011 [2]河北中医学院护理学院,河北石家庄050200

出　　处：《中华肿瘤防治杂志》2020年第23期1887-1895,共9页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金面上项目(81572441)。

摘　　要：目的探讨非受体型蛋白酪氨酸磷酸酶12(PTPN12)在胃贲门腺癌(GCA)的表达以及对胃癌细胞体外增殖、迁移和侵袭等生物学特性的影响。方法分别采用RT-qPCR和SP免疫组织化学法检测67例GCA组织及其相应癌旁正常组织中PTPN12mRNA和蛋白的表达情况,分析其与患者临床病理特征的关系;分别采用RT-qPCR和蛋白质印迹法方法检测PTPN12在胃癌细胞中mRNA和蛋白的表达;构建pcDNA3.1-PTPN12过表达质粒并转染BGC823细胞,检测其转染效率,并应用MTS法、克隆形成实验、划痕实验、Transwell小室分别检测转染前后细胞增殖、迁移和侵袭的变化。结果在GCA组织中,PTPN12mRNA的相对表达量显著低于其相应癌旁正常组织,Z=-10.359,P<0.01;与淋巴结转移(Z=-2.704,P=0.007)和TNM分期(Z=-3.699,P<0.01)相关。在GCA组织中,PTPN12蛋白阳性表达率与相应癌旁正常组织阳性表达率分别为32.84%(22/67)和79.10%(53/67),差异有统计学意义,χ2=29.101,P<0.01;并且与淋巴结转移(χ2=5.298,P=0.021)和TNM分期(χ2=5.761,P=0.016)相关;在胃癌细胞中PTPN12 mRNA和蛋白表达均低于对照组,F=862.733,P<0.01。在BGC823细胞中转染PTPN12过表达质粒,BGC823未转染组、pcDNA3.1-NC组和pcDNA3.1-PTPN12组PTPN12mRNA相对表达量分别为1.279±0.258、1.100±0.052和20.442±0.619,F=2457.892,P<0.01。BGC823未转染组与pcDNA3.1-NC组差异无统计学意义,t=1.178,P=0.304。转染后0、24、48、72和96h,BGC823未转染组细胞增殖速度分别为0.237±0.005、0.306±0.017、0.551±0.016、0.877±0.075和1.014±0.039,pcDNA3.1-NC组增殖速度分别为0.227±0.010、0.310±0.010、0.508±0.045、0.886±0.025和0.923±0.050,pcDNA3.1-PTPN12组增殖速度分别为0.232±0.010、0.261±0.010、0.395±0.025、0.676±0.035和0.678±0.038。在BGC823细胞中过表达PTPN12,与对照组相比,在72h其增殖能力显著减弱,F72h=22.668,P<0.01;96h其增殖能力显著减弱(F96h=66.812,P<0.01),克隆形成率显著减少(F=1484.140,P<0.01),抑制其侵�OBJECTIVE To investigate the expression of protein tyrosine phosphatase non-receptor type 12(PTPN12)in GCA and its effect on the biological characteristics of gastric cancer cells.METHODS RT-qPCR and SP immunohistochemistry(IHC)were used to detect the mRNA and protein of PTPN12in GCA tissues and corresponding normal tissues.Its relationship with clinicopathological features was analyzed.RT-qPCR and western blot were used to detect the expression of PTPN12mRNA and protein in gastric cancer cells.The pcDNA3.1-PTPN12over-expression plasmid was constructed and transfected in BGC823cells.The transfection efficiency was evaluated by RT-qPCR and western blot.The MTS,colony formation experiment,scratch test and Transwell chamber were used to detect the proliferation,migration and invasion of the cells before and after transfection.RESULTS In GCA tissues,the expression of PTPN12mRNA was obviously lower than that in corresponding normal tissues(Z=-10.359,P<0.001).It was also associated with lymph node metastasis(Z=-2.704,P=0.007)and TNM stage(Z=-3.699,P<0.001).In GCA tissues,the protein expression rate of PTPN12was obviously lower than that in corresponding normal tissues(χ2=29.101,P<0.001).It was closely correlated with lymph node metastasis(χ2=5.298,P=0.021),TNM stage(χ2=5.761,P=0.016);PTPN12mRNA and protein expression in gastric cancer cells were lower than that of control group(F=862.733,P<0.001).PTPN12 overexpression plasmid was transfected into BGC823cells,PTPN12expression was significantly higher than that of control group(F=2457.892,P<0.001).Overexpression of PTPN12in BGC823cells significantly decreased the proliferation ability(F72h=22.668,P<0.001;F96h=66.812,P<0.001).The colony formation rate was significantly reduced(F=1484.140,P<0.001),and the invasive ability was inhibited(F=234.999,P<0.001).CONCLUSIONS PTPN12is down-regulated in GCA and closely related to the development of GCA.Overexpression of PTPN12can inhibit the proliferation,migration and invasion of gastric cancer cells in vitro.

关 键 词：胃贲门腺癌 PTPN12 表达 生物学特性 

分 类 号：R735.2[医药卫生—肿瘤]