siRNA靶向抑制Tspan-1对结直肠癌细胞增殖和侵袭能力的影响
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摘要
目的探究siRNA靶向抑制Tspan-1对结直肠癌细胞增殖和侵袭能力的影响。方法实验组针对Tspan-1基因2个不同位点进行siRNA沉默,分别命名为Tspan-1-siRNA-1、Tspan-1-siRNA-2。采取完全剔除Tspan-1基因的作为阴性对照组,同时选取空白对照组。Wester blot方法检测各组的Tspan-1mRNA和蛋白的表达情况,选取基因沉默效果最强的一组进行结肠癌细胞HCT116细胞的转染,分别对转染前后细胞的增殖能力和侵袭能力进行检测,对siRNA靶向抑制Tspan-1能力进行评价和分析。结果相比于空白对照组,实验组各组细胞mRNA均出现了不同程度的下降,其中Tspan-1-siRNA-1、Tspan-1-siRNA-2,两组分别相比对照组下降61. 2%,57. 2%。随着转染时间的增加,实验组细胞增殖的速度明显放缓,其中第2、3、4、5天吸光度值要明显低于阴性对照组和空白对照组,差异有显著性(P <0. 05);同时两个对照组之间的吸光度值无显著差异(P> 0. 05);经过转染24 h后,实验组转移到膜上的细胞数要显著少于对照组,而其余两组无显著差异(P> 0. 05)。结论 Tspan-1基因经过siRNA沉默之后,能够有效降低结肠癌HCT116细胞的增殖和侵袭能力,siRNA靶向抑制Tspan-1基因可以作为治疗结直肠癌重要的的分子手段之一。
Objective To investigate the effect of siRNA targeting inhibition of Tspan-1 on proliferation and invasion of colorectal cancer cells. Methods Two loci of siRNA silencing of Tspan-1 were named as Tspan-1-siRNA-1 and Tspan-1-siRNA-2. The negative control group were without Tspan-1 gene. The blank control group were with untreated Tspan-1. Tspan-1 mRNA and protein were tested by Western blotting to select the best silenced gene for HCT116 cell transfection. Cell proliferation and invasion before and after transfection were detected and inhibition efficacy were evaluated. Results Tspan-1-siRNA-1 and Tspan-1-siRNA-2 decreased Tspan-1 mRNA by 61. 2% and 57. 2%respectively compared with blank control group. With the increase of transfection time,cells in experimental group showed lowered proliferation rate compared with two control groups. Cells in experimental group metastasized less than two control groups( P < 0. 05). Conclusion Tspan-1 gene silenced by siRNA can effectively inhibit proliferation and invasion of colon cancer cells and might be molecularly effective against colorectal cancer.
Objective To investigate the effect of siRNA targeting inhibition of Tspan-1 on proliferation and invasion of colorectal cancer cells. Methods Two loci of siRNA silencing of Tspan-1 were named as Tspan-1-siRNA-1 and Tspan-1-siRNA-2. The negative control group were without Tspan-1 gene. The blank control group were with untreated Tspan-1. Tspan-1 mRNA and protein were tested by Western blotting to select the best silenced gene for HCT116 cell transfection. Cell proliferation and invasion before and after transfection were detected and inhibition efficacy were evaluated. Results Tspan-1-siRNA-1 and Tspan-1-siRNA-2 decreased Tspan-1 mRNA by 61. 2% and 57. 2%respectively compared with blank control group. With the increase of transfection time,cells in experimental group showed lowered proliferation rate compared with two control groups. Cells in experimental group metastasized less than two control groups( P < 0. 05). Conclusion Tspan-1 gene silenced by siRNA can effectively inhibit proliferation and invasion of colon cancer cells and might be molecularly effective against colorectal cancer.
引文
[1]王惠芬.TSPAN-1和VEGF-C在宫颈鳞癌中的表达及意义[D].成都:四川医科大学,2015.
[2]南燕,李红雨,许雅娟.宫颈癌组织中TSPAN-1与Ki-67蛋白的表达[J].郑州大学学报(医学版),2010,45(3):469-471.
[3]李挺,徐美荣,王鼎,等.结肠癌中TSPAN-1和CORTACTIN蛋白的表达及其临床意义[J].中国普通外科杂志,2010,19(10):1107-1112.
[4]李挺,丛超,王星际,等.结直肠肿瘤组织中皮动蛋白的表达与临床病理意义及其与Arg和Tspan-1相关性的研究[J].中医临床研究,2015,7(33):131-133.
[5]何沛嵘.Tspan5在胃癌中的表达及调节胃癌细胞增殖机制的研究[D].广州:南方医科大学,2016.
[6]王惠芬,毛熙光.肿瘤增殖相关分子(TSPAN-1)和血管内皮生长因子C(VEGF-C)在宫颈鳞癌中的表达及意义[J].当代医学,2016,22(15):1-4.
[7]杨勇.HER2和TSPAN-1蛋白在胃癌组织中的表达及临床病理意义[D].遵义:遵义医学院,2016.
[8]南燕.宫颈鳞癌组织中TSPAN-1蛋白的表达及其与Ki-67和CD105相关性的研究[D].郑州:郑州大学,2010.
[9]周心一,金建强,齐晓薇.结直肠癌中CD166和TSPAN-1蛋白的表达及其临床意义[J].航空航天医学杂志,2013,24(11):1315-1318.
[10]Chen L,Zhu YY,Zhang XJ,et al.TSPAN1 protein expression:a significant prognostic indicator for patients with colorectal adenocarcinoma[J].World J Gastroenterol,2009,15(18):2270-2276.
[11]邢娜娜,张慧慧,谢佳琪,等.敲除TSPAN-1抑制人结肠癌细胞的生长和侵袭[J].医学研究杂志,2016,45(1):29-33.
[12]张泽栋,周逢强,徐宏,等.siRNA抑制TSPAN-1基因表达对结肠癌HCT116细胞株增殖和侵袭的影响[J].中国现代普通外科进展,2015,18(5):348-351.
[13]Detchokul S,Williams ED,Parker MW,et al.Tetraspanins asregulators of the tumour microenvironment:implications for metas-tasis and therapeutic strategies[J].Br J Pharmacol,2014,171(24):5462-5490.
[14]石刚,董明,盛伟伟,等.Tspan1及Integrinα6在人胰腺导管腺癌中的表达及其与临床特征的关系[J].中华外科杂志,2014,52(10):781-786.
[15]Lu Z,Luo T,Nie M,et al.TSPAN1 functions as an oncogene ingastric cancer and is downregulated by miR-573[J].FEBS Lett,2015,589(15):1988-1994.
[16]Xu F,Gao Y,Wang Y,et al.Decreased TSPAN1 promotes pros-tate cancer progression and is a marker for early biochemical recur-rence after radical prostatectomy[J].Oncotarget,2016,7(39):63294-63305.
[17]Hu T,Li Z,Gao CY,et al.Mechanisms of drug resistance in colon cancer and its therapeutic strategies[J].World J Gastroen-terol,2016,22(30):6876-6889.
[18]张志,吴亚运,何宋兵,等.257例大肠癌患者的临床特征、病理及预后分析[J].中国血液流变学杂志,2016,26(1):76-78.
[2]南燕,李红雨,许雅娟.宫颈癌组织中TSPAN-1与Ki-67蛋白的表达[J].郑州大学学报(医学版),2010,45(3):469-471.
[3]李挺,徐美荣,王鼎,等.结肠癌中TSPAN-1和CORTACTIN蛋白的表达及其临床意义[J].中国普通外科杂志,2010,19(10):1107-1112.
[4]李挺,丛超,王星际,等.结直肠肿瘤组织中皮动蛋白的表达与临床病理意义及其与Arg和Tspan-1相关性的研究[J].中医临床研究,2015,7(33):131-133.
[5]何沛嵘.Tspan5在胃癌中的表达及调节胃癌细胞增殖机制的研究[D].广州:南方医科大学,2016.
[6]王惠芬,毛熙光.肿瘤增殖相关分子(TSPAN-1)和血管内皮生长因子C(VEGF-C)在宫颈鳞癌中的表达及意义[J].当代医学,2016,22(15):1-4.
[7]杨勇.HER2和TSPAN-1蛋白在胃癌组织中的表达及临床病理意义[D].遵义:遵义医学院,2016.
[8]南燕.宫颈鳞癌组织中TSPAN-1蛋白的表达及其与Ki-67和CD105相关性的研究[D].郑州:郑州大学,2010.
[9]周心一,金建强,齐晓薇.结直肠癌中CD166和TSPAN-1蛋白的表达及其临床意义[J].航空航天医学杂志,2013,24(11):1315-1318.
[10]Chen L,Zhu YY,Zhang XJ,et al.TSPAN1 protein expression:a significant prognostic indicator for patients with colorectal adenocarcinoma[J].World J Gastroenterol,2009,15(18):2270-2276.
[11]邢娜娜,张慧慧,谢佳琪,等.敲除TSPAN-1抑制人结肠癌细胞的生长和侵袭[J].医学研究杂志,2016,45(1):29-33.
[12]张泽栋,周逢强,徐宏,等.siRNA抑制TSPAN-1基因表达对结肠癌HCT116细胞株增殖和侵袭的影响[J].中国现代普通外科进展,2015,18(5):348-351.
[13]Detchokul S,Williams ED,Parker MW,et al.Tetraspanins asregulators of the tumour microenvironment:implications for metas-tasis and therapeutic strategies[J].Br J Pharmacol,2014,171(24):5462-5490.
[14]石刚,董明,盛伟伟,等.Tspan1及Integrinα6在人胰腺导管腺癌中的表达及其与临床特征的关系[J].中华外科杂志,2014,52(10):781-786.
[15]Lu Z,Luo T,Nie M,et al.TSPAN1 functions as an oncogene ingastric cancer and is downregulated by miR-573[J].FEBS Lett,2015,589(15):1988-1994.
[16]Xu F,Gao Y,Wang Y,et al.Decreased TSPAN1 promotes pros-tate cancer progression and is a marker for early biochemical recur-rence after radical prostatectomy[J].Oncotarget,2016,7(39):63294-63305.
[17]Hu T,Li Z,Gao CY,et al.Mechanisms of drug resistance in colon cancer and its therapeutic strategies[J].World J Gastroen-terol,2016,22(30):6876-6889.
[18]张志,吴亚运,何宋兵,等.257例大肠癌患者的临床特征、病理及预后分析[J].中国血液流变学杂志,2016,26(1):76-78.