Tectonic家族成员1对T47D细胞增殖和细胞周期的影响  

Influence of down-regulation of Tectonic family member 1 on proliferation and cycle of T47D cells

作　　者：梁冰[1] 梁晓丹 裴新红[2] LIANG Bing;LIANG Xiao-dan;PEI Xin-hong(Department of Breast Surgery,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450003,China;Department of Breast Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450052,China)

机构地区：[1]郑州大学第二附属医院乳腺外科,河南郑州450003 [2]郑州大学第一附属医院乳腺外科,河南郑州450052

出　　处：《中华实用诊断与治疗杂志》2022年第7期686-689,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河南省高等学校重点科研项目(22A320004)。

摘　　要：目的观察乳腺癌细胞Tectonic家族成员1(Tectonic family member 1,TCTN1)表达变化,探讨下调其表达对T47D细胞增殖及细胞周期的影响。方法取对数生长期MCF-7、MDA-MB-231、T47D细胞及正常乳腺MCF-10A细胞,采用实时荧光定量PCR法检测TCTN1 mRNA相对表达量,筛选出TCTN1 mRNA相对表达量最高的T47D细胞进行后续试验。取对数生长期T47D细胞,分为转染组(转染siRNA-TCTN1)、阴性对照组(转染siRNA-NC)和空白对照组(不作处理)。转染48 h,采用实时荧光定量PCR法检测3组TCTN1 mRNA相对表达量,采用Western blot法检测TCTN1蛋白相对表达量,采用流式细胞术检测细胞周期;转染后培养0、24、48、72、96 h,采用CCK-8法检测细胞增殖率。结果MCF-7、MDA-MB-231、T47D细胞TCTN1 mRNA相对表达量(3.120±0.893、2.839±1.102、5.932±0.938)均高于MCF-10A细胞(1.023±0.182)(P<0.05),T47D细胞TCTN1 mRNA相对表达量高于MCF-7、MDA-MB-231细胞(P<0.05)。转染48 h,转染组TCTN1 mRNA及蛋白相对表达量(0.539±0.021、0.050±0.003)、G/G期细胞比率[(46.06±1.30)%]均低于阴性对照组[0.932±0.102、0.379±0.052、(54.67±1.13)%]和空白对照组[1.021±0.091、0.457±0.017、(53.77±1.91)%](P<0.05),G/M期细胞比率[(22.49±1.45)%]高于阴性对照组[(14.60±2.44)%]和空白对照组[(14.99±2.16)%](P<0.05),阴性对照组与空白对照组比较差异均无统计学意义(P>0.05)。转染后培养0、24 h,3组细胞增殖率比较差异均无统计学意义(P>0.05);转染后培养48、72、96 h,转染组细胞增殖率[(53.0±8.2)%、(78.0±10.2)%、(123.0±11.2)%]低于阴性对照组[(83.0±13.5)%、(129.0±13.9)%、(172.5±28.3)%]和空白对照组[(92.0±14.7)%、(139.2±19.3)%、(192.3±29.2)%](P<0.05),阴性对照组与空白对照组比较差异无统计学意义(P>0.05)。结论MCF-7、MDA-MB-231、T47D细胞TCTN1 mRNA表达上调;下调TCTN1表达可抑制T47D细胞增殖,使细胞G/M期阻滞。Objective To observe the changes of Tectonic family member 1(TCTN1)in breast cancer cells,and to explore the influence of down-regulating its expression on the proliferation and cell cycle of T47Dcells.Methods The MCF-7,MDA-MB-231and T47Dcells in logarithmic growth phase and the normal breast MCF-10Acells were selected,and the relative expression of TCTN1mRNA was detected by real-time fluorescence quantitative PCR.The T47Dcells with the highest relative expression of TCTN1mRNA was screened out for subsequent experiments.The T47Dcells in logarithmic growth phase were divided into transfection group(transfected with siRNA-TCTN1),negative control group(transfected with siRNA-NC)and blank control group(no treatment).After 48-h transfection,the relative expression of TCTN1mRNA was detected in three groups by real-time fluorescence quantitative PCR,the relative expression of TCTN1protein was detected by Western blot,and the cell cycle was detected by flow cytometry.The proliferation rate was detected by CCK-8 method after culture for 0,24,48,72and 96h.Results The relative expression of TCTN1mRNA was significantly higher in MCF-7cells(3.120±0.893),MDA-MB-231cells(2.839±1.102)and T47Dcells(5.932±0.938)than that in MCF-10Acells(1.023±0.182)(P<0.05),and higher in T47Dcells than that in MCF-7cells and MDA-MB-231cells(P<0.05).After 48-h transfection,the relative expressions of TCTN1mRNA and protein as well as the G/Gcell ratio were significantly lower in transfection group[0.539±0.021,0.050±0.003,(46.06±1.30)%]than those in negative control group[0.932±0.102,0.379±0.052,(54.67±1.13)%]and blank control group[1.021±0.091,0.457±0.017,(53.77±1.91)%](P<0.05),the G/M cell ratio was higher in transfection group[(22.49±1.45)%]than that in negative control group[(14.60±2.44)%]and blank control group[(14.99±2.16)%](P<0.05),and all above indexes showed no significant differences between negative control group and blank control group(P>0.05).After 0and 24-h transfection,the proliferation rates showed no significant diffe

关 键 词：乳腺癌 Tectonic家族成员1 T47D细胞 细胞增殖 细胞周期 

分 类 号：R737.9[医药卫生—肿瘤]