脂联素分子受体3在抑制转化生长因子-β介导的食管-胃交界腺癌转移中的功能

The function of progestin and adipoQ receptor 3 in the inhibition of transforming growth factor-mediated metastasis of esophageal-gastric junction adenocarcinoma at animal level

目的探讨脂联素分子受体3(PAQR3)在抑制转化生长因子-β(TGF-β)介导的食管-胃交界腺癌转移中的功能。方法选用OE19细胞系和30只BALB/c无特定病原体(SPF)级裸鼠进行实验, 随机分成3组:A组(对照组)、B组(TGF-β通路激活组)及C组(TGF-β通路激活的同时过表达PAQR3), 每组各10只裸鼠, 建立裸鼠皮下移植瘤模型, 观察过表达PAQR3对TGF-β介导的OE19细胞皮下成瘤及转移能力的影响;采用定量RT-PCR检测裸鼠肿瘤组织中p53、细胞核增殖抗原(Ki-67)、TGF-β信号通路及上皮-间充质转化(EMT)相关基因的mRNA表达水平, 组间比较采用单因素方差分析。结果 A组10只裸鼠中6只成瘤, 其中4只发生转移;B组10只裸鼠中8只成瘤、6只发生转移;C组10只裸鼠中3只成瘤、均未发生转移。B组动物肿瘤组织中TGF-β、Ki-67、p53、Twist、Snail、Vimentin mRNA表达量(13.52±0.07、5.23±0.04、2.34±0.02、13.74±0.07、8.23±0.05、7.73±0.05)均明显高于A组(1.00±0.01、1.00±0.02、1.00±0.01、1.00±0.01、1.00±0.02、1.00±0.02, t=32.825、17.682、3.901、33.834、27.612、24.558, 均P<0.01);B组动物肿瘤组织中PAQR3、E-cadherin mRNA表达量(0.41±0.03、0.31±0.03)均明显低于A组(1.00±0.02、1.00±0.02, t=8.735, P<0.05, t=10.846, P<0.01)。C组动物肿瘤组织中TGF-β、PAQR3、E-cadherin mRNA表达量(13.23±0.08、5.22±0.04、2.93±0.03)均明显高于A组(1.00±0.01、1.00±0.02、1.00±0.02, t=31.752、17.554、4.045, 均P<0.01);C组动物肿瘤组织中Ki-67、p53、Twist、Snail、Vimentin mRNA表达量(0.31±0.03、0.40±0.05、0.24±0.03、0.22±0.03、0.26±0.04)均明显低于A组(1.00±0.02, t=11.356, P<0.01;1.00±0.01, t=9.558, P<0.05;1.00±0.01, t=12.391, P<0.01;1.00±0.02, t=12.845, P<0.01;1.00±0.02, t=11.034, P<0.01)。结论 PAQR3作为新型抑癌基因, 具有在体内抑制TGF-β介导的食管-胃交界腺癌细胞皮下成瘤及转移中的功能。

Objective To explore the function of progestin and adipoQ receptor 3(PAQR3)in inhibiting transforming growth factor(TGF)-mediated metastasis of esophageal-gastric junction adenocarcinoma(EJA)at animal level.Methods Using the OE19 cell line and 30 BALB/c SPF grade nude mice,the experiments were randomly divided into three groups:group A(control group),group B(TGF-βpathway activation group)and group C(simultaneous overexpression of PAQR3 with TGF-βpathway activation),10 nude mice were each in each group,then observe the effect of PAQR3 on TGF-βmediated subcutaneous tumorigenesis and metastatic ability,and apply quantitative RT-PCR to analyze the mRNA expression of p53,proliferation cell nuclear antigen(Ki-67),TGF-βsignaling related genes,and epithelial-mesenchymal transition(EMT)related genes in animal tumor tissues.Statistical analysis was performed using SPSS 19.0 statistical software,and the inter-group comparisons were performed using a one-way ANOVA.Results The subcutaneous neoplasia was found in six of ten nude mice in group A,four of whom had metastases;the subcutaneous neoplasia was found in eight of ten nude mice in group B,six of whom had metastases;the subcutaneous neoplasia was found in three of ten nude mice in group C,none had metastasis.The expression of TGF-β,Ki-67,p53,Twist,Snail and Vimentin in animal tumor tissues of B group(13.52±0.07,5.23±0.04,2.34±0.02,13.74±0.07,8.23±0.05,7.73±0.05)were significantly higher than those in the control group(A group)(1.00±0.01,1.00±0.02,1.00±0.01,1.00±0.01,1.00±0.02,1.00±0.02;t=32.825,17.682,3.901,33.834,27.612,24.558;all P<0.01).The expression of PAQR3 and E-cadherin mRNA in animal tumor tissues of B group(0.41±0.03、0.31±0.03)were significantly lower than those in the control group(A group)(1.00±0.02,1.00±0.02;t=8.735,P<0.05;t=10.846,P<0.01).The expression of TGF-β,PAQR3 and E-cadherin mRNA in animal tumor tissues of C group(13.23±0.08,5.22±0.04,2.93±0.03)were significantly higher than those in the control group(A group)(1.00±0.01,1.00±0.02,1.00±0.02;t=31.752,17.554,4.045;all P<0.01).The expression of Ki-67,p53,Twist,Snail and Vimentin mRNA in animal tumor tissues of C group(0.31±0.03,0.40±0.05,0.24±0.03,0.22±0.03,0.26±0.04)were significantly lower than those in the control group(A group)(1.00±0.02,t=11.356,P<0.01;1.00±0.01,t=9.558,P<0.05;1.00±0.01,t=12.391,P<0.01;1.00±0.02,t=12.845,P<0.01;1.00±0.02,t=11.034,P<0.01).Conclusion As a novel tumor suppressor gene,PAQR3 has a function in inhibiting TGF-βmediated subcutaneous subcutaneous tumorigenesis and metastasis of esophageal-gastric junction adenocarcinoma in vivo.