过表达STAT1与CD74CD44促进结肠癌细胞粘附和迁移的相关性
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摘要
目的探讨信号转导和转录激活子1(STAT1)与CD74/CD44在结肠癌中异常表达与肿瘤转移的相关性。方法培养结肠癌细胞株Ls-174-T,待其生长至对数生长期,传至第3代收集肿瘤细胞,各细胞分为3份。1份行RTPCR、Western blotting方法检测STAT1与CD74/CD44表达情况;1份检测定量粘附细胞数;1份以划痕实验检测细胞迁移能力。统计分析比较STAT1与CD74/CD44在结肠癌细胞中的异常表达情况及与肿瘤迁移和粘附能力的相关性。结果RT-PCR、Western blotting结肠癌组织中STAT1与CD74/CD44的蛋白及mRNA表达显著高于正常结肠组织,两组相比具有统计学差异(P<0.05)。MTT法检测结果显示,不同作用时间(0、6、12、24、48 h)不同剂量STAT1与CD74/CD44过表达质粒(0.000、0.001、0.010、0.060、0.100 mg/ml)干预后,结肠癌细胞体外粘附能力明显加强,与对照组(0.000 mg/ml各组)相比,差异有统计学意义;Transwell结果显示,不同作用时间(0、6、12、24、48 h)不同剂量STAT1与CD74/CD44过表达质粒(0.000、0.001、0.010、0.060、0.100 mg/ml)干预后,结肠癌细胞体外迁移能力随着时间及剂量的增加而明显加强,与对照组(0.000mg/ml各组)相比,差异有统计学意义。结论 STAT1与CD74/CD44在结肠癌细胞中高表达,从而调节结肠癌细胞粘附和迁移能力的作用,表明STAT1与CD74/CD44在结肠癌发生进展中可能与肿瘤的侵袭和转移密切相关。
Objective To investigate the signal transducer and activator of transcription 1(STAT1) and CD74/CD44 expression in colon cancer cells with abnormal tumor metastasis. Methods Cultured colon cancer cell line Ls-174-T,let it grow to a logarithmic growth phase,the third passage collected tumor cells,each cell was divided into 3 parts. A line of RT-PCR,Western blotting method was used to detect STAT1 and CD74/CD44 expression;a quantitative detection of adherent cells;a scratch test to detect cell migration. Statistical analysis and comparison of STAT1 and CD74/CD44 in colon cancer cells and abnormal expression correlated with tumor migration and adhesion ability. Results The expression of STAT1 and CD74/CD44 protein in colorectal cancer tissues was significantly higher than that in normal colorectal tissues( P < 0. 05). MTT assay test results showed that different time(0,6,12,24,48 h) and different doses of STAT1 and CD74/CD44 over-expression plasmid(0. 000,0. 001,0. 010,0. 060,0. 100 mg/ml) intervention colon cancer cells In vitro adhesion significantly strengthened with the control group(0. 000 mg/ml in each group),the difference was statistically significant;Transwell results showed that different time(0,6,12,24,48 h) and different doses of STAT1 CD74/CD44 over-expression plasmid(0. 000,0. 001,0. 010,0. 060,0. 100 mg/ml) after the intervention of colon cancer cells in vitro migration with increasing dose and time and significantly strengthened with the control group(0. 000 mg/ml in each group),the difference was statistically significant. Conclusion STAT1 and CD74/CD44 is highly expressed in colon cancer cells,thereby regulating the role of colon cancer cell adhesion and migration,indicating that STAT1 and CD74/CD44 may be associated with tumor invasion and metastasis in colon cancer progression.
Objective To investigate the signal transducer and activator of transcription 1(STAT1) and CD74/CD44 expression in colon cancer cells with abnormal tumor metastasis. Methods Cultured colon cancer cell line Ls-174-T,let it grow to a logarithmic growth phase,the third passage collected tumor cells,each cell was divided into 3 parts. A line of RT-PCR,Western blotting method was used to detect STAT1 and CD74/CD44 expression;a quantitative detection of adherent cells;a scratch test to detect cell migration. Statistical analysis and comparison of STAT1 and CD74/CD44 in colon cancer cells and abnormal expression correlated with tumor migration and adhesion ability. Results The expression of STAT1 and CD74/CD44 protein in colorectal cancer tissues was significantly higher than that in normal colorectal tissues( P < 0. 05). MTT assay test results showed that different time(0,6,12,24,48 h) and different doses of STAT1 and CD74/CD44 over-expression plasmid(0. 000,0. 001,0. 010,0. 060,0. 100 mg/ml) intervention colon cancer cells In vitro adhesion significantly strengthened with the control group(0. 000 mg/ml in each group),the difference was statistically significant;Transwell results showed that different time(0,6,12,24,48 h) and different doses of STAT1 CD74/CD44 over-expression plasmid(0. 000,0. 001,0. 010,0. 060,0. 100 mg/ml) after the intervention of colon cancer cells in vitro migration with increasing dose and time and significantly strengthened with the control group(0. 000 mg/ml in each group),the difference was statistically significant. Conclusion STAT1 and CD74/CD44 is highly expressed in colon cancer cells,thereby regulating the role of colon cancer cell adhesion and migration,indicating that STAT1 and CD74/CD44 may be associated with tumor invasion and metastasis in colon cancer progression.
引文
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[3]Merlos-Suárez A,Barriga FM,Jung P,et al.The intestinal stem cell signature identifies colorectal cancer stem cells and predicts disease relapse[J].Cell Stem Cell,2011,8(5):511-524.
[4]Salazar R,Roepman P,Capella G,et al.Gene expression signature to improve prognosis prediction of stageⅡandⅢcolorectal cancer[J].J Clin Oncol,2011,29(1):17-24.
[5]Oh SC,Park YY,Park ES,et al.Prognostic gene expression signature associated with two molecularly distinct subtypes of colorectal cancer[J].Gut,2012,61(2):1291-1298.
[6]Mettu RK,Wan YW,Habermann JK.A 12-gene genomic instability signature predicts clinical outcomes in multiple cancer types[J].Int J Biol Markers,2010,25(4):219-228.
[7]Peng J,Wang Z,Chen W,et al.Integration of genetic signature and TNM staging system for predicting the relapse of locally advanced colorectal cancer[J].Int J Colorectal Dis,2010,25(11):1277-1285.
[8]Kennedy RD,Bylesjo M,Kerr P,et al.Development and independent validation of a prognostic assay for stageⅡcolon cancer using formalin-fixed paraffin-embedded tissue[J].J Clin Oncol,2011,29(35):4620-4626.
[9]Sveen A,Gesen TH,Nesbakken A,et al.Colo Guide Pro:a prognostic 7-gene expression signature for stageⅢcolorectal cancer patients[J].Clin Cancer Res,2012,18(21):6001-6010.
[10]Tsuji S,Midorikawa Y,Takahashi T,et al.Potential responders to FOLFOX therapy for colorectal cancer by Random Forests analysis[J].Br J Cancer,2012,106(1):126-132.
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[12]Merlos-Suarez A,Barriga FM,Jung P,et al.The intestinal stem cell signature identifies colorectal cancer stem cells and predicts disease relapse[J].Cell Stem Cell,2011,8(5):511-524.
[13]Aceto N,Sausgruber N,Brinkhaus H,et al.Tyrosine phosphatase SHP2 promotes breast cancer progression and maintains tumor-initiating cells via activation of key transcription factors and a positive feedback signaling loop[J].Nat Med,2012,18(4):529-537.
[14]Biagi JJ,Raphael MJ,Mackillop WJ,et al.Association between time to initiation of adjuvant chemotherapy and survival in colorectal cancer:a systematic review and meta-analysis[J].JAMA,2011,305(22):2335-2342.