特异AT序列结合蛋白2对肾癌细胞生物学行为的影响  

Effects of special AT-rich sequence-binding protein 2 on biological behavior of renal cell carcinoma

作　　者：赵鹏程 杨栋 何朝红 田沛 Zhao Pengcheng;Yang Dong;He Chaohong;Tian Pei(Department of Urology,the Affiliated Cancer Hospital of Zhengzhou University,Zhengzhou 450008,China)

机构地区：[1]郑州大学附属肿瘤医院泌尿外科,郑州450008

出　　处：《中华实验外科杂志》2024年第6期1152-1155,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划(LHGJ20220192)。

摘　　要：目的探讨特异AT序列结合蛋白2(SATB2)在肾癌细胞中的表达及其对肾癌细胞生物学行为的影响。方法选取人肾皮质近曲小管上皮细胞HK-2和肾癌细胞786-O、ACHN,荧光定量聚合酶链反应(PCR)法检测HK-2、786-O和ACHN细胞SATB2 mRNA的表达水平;将重组过表达质粒pcDNA3.1-SATB2及空载质粒pcDNA3.1-NC分别转染至786-O肾癌细胞中,采用细胞计数试剂盒(CCK-8)检测两组细胞增殖活力;采用Transwell实验检测两组细胞迁移与侵袭水平;采用蛋白质印迹法(Western blot)检测两组细胞SATB2及N-钙黏蛋白(N-cadherin)、E-钙黏蛋白(E-cadherin)、抗波形蛋白(Vimentin)的表达。组间比较采用t检验。结果786-O细胞(0.50±0.06)和ACHN细胞(0.57±0.07)中SATB2 mRNA的表达水平明显低于HK-2细胞(1.54±0.10),差异有统计学意义(t=23.13、20.23,P<0.05)。pcDNA3.1-SATB2细胞(0.24±0.02)和pcDNA3.1-NC细胞(0.28±0.03)在第24小时的吸光度(A)值差异无统计学意义(t=2.06,P>0.05),而在第48、72小时,pcDNA3.1-SATB2细胞(0.52±0.03、0.80±0.05)的A值明显低于pcDNA3.1-NC细胞(0.69±0.04、1.11±0.08),差异有统计学意义(t=7.67、8.22,P<0.05)。pcDNA3.1-SATB2细胞迁移细胞数[(76.83±6.82)个]和侵袭细胞数[(53.33±5.35)个]明显低于pcDNA3.1-NC细胞[(135.83±7.96)、(112.67±5.82)个],差异有统计学意义(t=13.78、18.38,P<0.05)。pcDNA3.1-SATB2细胞SATB2(2.01±0.12)、E-cadherin(1.53±0.08)蛋白表达水平明显高于pcDNA3.1-NC细胞(0.38±0.03、0.50±0.05),差异有统计学意义(t=31.82、27.76,P<0.05)。pcDNA3.1-SATB2细胞N-cadherin(0.86±0.05)、Vimentin(0.66±0.05)蛋白表达水平明显低于pcDNA3.1-NC细胞(1.66±0.08、1.29±0.05),差异有统计学意义(t=20.16、20.84,P<0.05)。结论SATB2在肾癌细胞中表达下降,过表达SATB2可通过调控肾癌细胞的上皮-间充质转化抑制肿瘤细胞的迁移和侵袭。Objective To investigate the effects of special AT-rich sequence-binding protein 2(SATB2)on biological behavior of renal cell carcinoma(RCC).Methods Human renal cortex proximal convoluted tubule epithelial cells(HK-2)and RCC cells(786-O and ACHN)were selected.The expression level of SATB2 mRNA in HK-2,786-O and ACHN cells was detected by quantitative real-time polymerase chain reaction(PCR).Recombinant overexpressed plasmid pcDNA3.1-SATB2 and empty plasmid pcDNA3.1-NC were transfected into 786-O cells,respectively.The cell counting kit-8(CCK-8)was used to detect cell proliferation activity in the two groups.The abilities of migration and invasion were detected by Transwell assay.Western blotting(WB)was used to detect the expression of SATB2,N-cadherin,E-cadherin and Vimentin in the two groups.The inter group comparison was conducted using t-test.Results The expression level of SATB2 mRNA in 786-O cells(0.50±0.06)and ACHN cells(0.57±0.07)was significantly lower than that in HK-2 cells(1.54±0.10,t=23.13,20.23,P<0.05).The absorbance(A)values of pcDNA3.1-SATB2 cells(0.24±0.02)and pcDNA3.1-NC cells(0.28±0.03)at 24 h were not significantly different(t=2.06,P>0.05).The A value of pcDNA3.1-SATB2 cells(0.52±0.03,0.80±0.05)at 48 h and 72 h was significantly lower than that of pcDNA3.1-NC cells[(0.69±0.04),(1.11±0.08),t=7.67,8.22,P<0.05].The number of migrating cells(76.83±6.82)and invading cells(53.33±5.35)of pcDNA3.1-SATB2 cells was significantly less than that of pcDNA3.1-NC cells(135.83±7.96,112.67±5.82;t=13.78,18.38,P<0.05).The expression levels of SATB2(2.01±0.12)and E-cadherin(1.53±0.08)in pcDNA3.1-SATB2 cells were significantly higher than those in pcDNA3.1-NC cells[(0.38±0.03),(0.50±0.05),t=31.82,27.76,P<0.05].The expression levels of N-cadherin(0.86±0.05)and Vimentin(0.66±0.05)in pcDNA3.1-SATB2 cells were significantly lower than those in pcDNA3.1-NC cells[(1.66±0.08),(1.29±0.05),t=20.16,20.84,P<0.05].Conclusion SATB2 is low expressed in RCC,and overexpression of SATB2 can inhibit the migratio

关 键 词：肾细胞癌 特异AT序列结合蛋白2 上皮-间充质转化 迁移 侵袭 

分 类 号：R737.11[医药卫生—肿瘤]