肺癌细胞中SOX1调控ECT2的表达
|
摘要
目的探讨肺癌细胞中SOX1基因是否直接调控ECT2的表达。方法运用甲基化特异PCR法检测50例原发肺癌和癌旁组织中SOX1的甲基化水平。以肺癌细胞株A-1细胞为研究对象,Realtime-PCR检测ECT2表达情况。通过荧光素酶报告检查法以及染色体免疫共沉淀方法检测SOX1基因是否直接调控ECT2的表达。免疫组化检测50例肺癌组织中ECT2与SOX1表达的关系。结果原发性肺癌的甲基化异常检出率是70%(35/50)。过表达SOX1后A-1细胞中ECT2表达下降,而si RNA抑制SOX1后ECT2表达升高,荧光素酶报告检查法以及染色体免疫共沉淀方法提示SOX1可直接与ECT2启动子结合,抑制ECT2的表达。SOX1与ECT2在肺癌组织中表达负相关(r=-0.433,P=0.001)。结论肺癌细胞A-1中SOX1基因直接调控ECT2的表达。
Objective To test whether SOX1 is a novel transcriptional repressor for ECT2 in human lung cancer. Methods 50 primary lung cancer tissues and corresponding normal tissues were detected by Methylation-specific PCR method.The expression of ECT2 in the A-1 cell was detected by Realtime-PCR.We determined whether SOX1 may regulate the expression of ECT2 by using Chromin immunoprecipitation(Ch IP) assay and Luciferase activity assay.Immunohistochemical analysis was used to test whether there is an inverse correlation between ECT2 and SOX1. Results Aberrant methylation in primary lung cancer was 70%(35/50).Ectopic expression of SOX1 in the A-1 cell repressed ECT2 expression.Conversely,si RNA silencing of the SOX1 gene increased ECT2 expression.We also show that SOX1 directly interacted with and repressed the ECT2 promoter.Moreover,the analysis of 50 primary lung cancer samples revealed an inverse correlation between ECT2 and SOX1 levels(r=-0.433,P=0.001). Conclusion SOX1 is a novel transcriptional repressor for ECT2 in human lung cancer.
Objective To test whether SOX1 is a novel transcriptional repressor for ECT2 in human lung cancer. Methods 50 primary lung cancer tissues and corresponding normal tissues were detected by Methylation-specific PCR method.The expression of ECT2 in the A-1 cell was detected by Realtime-PCR.We determined whether SOX1 may regulate the expression of ECT2 by using Chromin immunoprecipitation(Ch IP) assay and Luciferase activity assay.Immunohistochemical analysis was used to test whether there is an inverse correlation between ECT2 and SOX1. Results Aberrant methylation in primary lung cancer was 70%(35/50).Ectopic expression of SOX1 in the A-1 cell repressed ECT2 expression.Conversely,si RNA silencing of the SOX1 gene increased ECT2 expression.We also show that SOX1 directly interacted with and repressed the ECT2 promoter.Moreover,the analysis of 50 primary lung cancer samples revealed an inverse correlation between ECT2 and SOX1 levels(r=-0.433,P=0.001). Conclusion SOX1 is a novel transcriptional repressor for ECT2 in human lung cancer.
引文
[1]Boelens MC,Kok K,Van der Vlies P,et al.Genomic aberrations in squamous cell lung carcinoma related to lymph node or distant metastasis[J].Lung cancer(Amsterdam,Netherlands),2009,66(3):372-378.
[2]Vural B,Chen LC,Saip P,et al.Frequency of SOX Group B(SOX1,2,3)and ZIC2 antibodies in Turkish patients with small cell lung carcinoma and their correlation with clinical parameters[J].Cancer,2005,103(12):2575-2583.
[3]Tsao CM,Yan MD,Shih YL,et al.SOX1 functions as a tumor suppressor by antagonizing the WNT/betacatenin signaling pathway in hepatocellular carcinoma[J].Hepatology(Baltimore,Md),2012,56(6):2277-2287.
[4]Guan Z,Zhang J,Wang J,et al.SOX1 Down-regulates beta-catenin and Reverses Malignant Phenotype in Nasopharyngeal Carcinoma[J].Molecular Cancer,2014,13(1):257.
[5]Li N,Li X,Li S,et al.Cisplatin-induced downregulation of SOX1 increases drug resistance by activating autophagy in non-small cell lung cancer cell[J].Biochemical and Biophysical Research Communications,2013,439(2):187-190.
[6]Shih YL,Hsieh CB,Yan MD,et al.Frequent concomitant epigenetic silencing of SOX1 and secreted frizzled-related proteins(SFRPs)in human hepatocellular carcinoma[J].Journal of Gastroenterology and Hepatology,2013,28(3):551-559.
[7]Guan Z,Zhang J,Wang J,et al.SOX1 down-regulates betacatenin and reverses malignant phenotype in nasopharyngeal carcinoma[J].Molecular Cancer,2014,13:257.
[8]Wentzensen N,Bakkum-Gamez JN,Killian JK,et al.Discovery and validation of methylation markers for endometrial cancer[J].International Journal of Cancer,2014,135(8):1860-1868.
[9]Ji XD,Li G,Feng YX,et al.Eph B3 is overexpressed in non-small-cell lung cancer and promotes tumor metastasis by enhancing cell survival and migration[J].Cancer Research,2011,71(3):1156-1166.
[10]Kan L,Israsena N,Zhang Z,et al.Sox1 acts through multiple independent pathways to promote neurogenesis[J].Developmental Biology,2004,269(2):580-594.
[11]Akiyama H,Lyons JP,Mori-Akiyama Y,et al.Interactions between Sox9 and beta-catenin control chondrocyte differentiation[J].Genes&Development,2004,18(9):1072-1087.
[12]Lin YW,Tsao CM,Yu PN,et al.SOX1 suppresses cell growth and invasion in cervical cancer[J].Gynecologic Oncology,2013,131(1):174-181.
[13]Lv Z,Hu M,Zhen J,et al.Rac1/PAK1 signaling promotes epithelial-mesenchymal transition of podocytes in vitro via triggering beta-catenin transcriptional activity under high glucose conditions[J].The International Journal of Biochemistry&Cell Biology,2013,45(2):255-264.
[14]Patel S,Takagi KI,Suzuki J,et al.RhoG TPase activation is a key step in renal epithelial mesenchymal transdifferentiation[J].J Am Soc Nephrol,2005,16(7):1977-1984.
[15]Liang Y,He L,Yuan H,et al.Association between RUNX3 promoter methylation and non-small cell lung cancer:a meta-analysis[J].Journal of Thoracic Disease,2014,6(6):694-705.
[16]Murata Y,Minami Y,Iwakawa R,et al.ECT2amplification and overexpression as a new prognostic biomarker for early-stage lung adenocarcinoma[J].Cancer Science,2014,105(4):490-497.
[2]Vural B,Chen LC,Saip P,et al.Frequency of SOX Group B(SOX1,2,3)and ZIC2 antibodies in Turkish patients with small cell lung carcinoma and their correlation with clinical parameters[J].Cancer,2005,103(12):2575-2583.
[3]Tsao CM,Yan MD,Shih YL,et al.SOX1 functions as a tumor suppressor by antagonizing the WNT/betacatenin signaling pathway in hepatocellular carcinoma[J].Hepatology(Baltimore,Md),2012,56(6):2277-2287.
[4]Guan Z,Zhang J,Wang J,et al.SOX1 Down-regulates beta-catenin and Reverses Malignant Phenotype in Nasopharyngeal Carcinoma[J].Molecular Cancer,2014,13(1):257.
[5]Li N,Li X,Li S,et al.Cisplatin-induced downregulation of SOX1 increases drug resistance by activating autophagy in non-small cell lung cancer cell[J].Biochemical and Biophysical Research Communications,2013,439(2):187-190.
[6]Shih YL,Hsieh CB,Yan MD,et al.Frequent concomitant epigenetic silencing of SOX1 and secreted frizzled-related proteins(SFRPs)in human hepatocellular carcinoma[J].Journal of Gastroenterology and Hepatology,2013,28(3):551-559.
[7]Guan Z,Zhang J,Wang J,et al.SOX1 down-regulates betacatenin and reverses malignant phenotype in nasopharyngeal carcinoma[J].Molecular Cancer,2014,13:257.
[8]Wentzensen N,Bakkum-Gamez JN,Killian JK,et al.Discovery and validation of methylation markers for endometrial cancer[J].International Journal of Cancer,2014,135(8):1860-1868.
[9]Ji XD,Li G,Feng YX,et al.Eph B3 is overexpressed in non-small-cell lung cancer and promotes tumor metastasis by enhancing cell survival and migration[J].Cancer Research,2011,71(3):1156-1166.
[10]Kan L,Israsena N,Zhang Z,et al.Sox1 acts through multiple independent pathways to promote neurogenesis[J].Developmental Biology,2004,269(2):580-594.
[11]Akiyama H,Lyons JP,Mori-Akiyama Y,et al.Interactions between Sox9 and beta-catenin control chondrocyte differentiation[J].Genes&Development,2004,18(9):1072-1087.
[12]Lin YW,Tsao CM,Yu PN,et al.SOX1 suppresses cell growth and invasion in cervical cancer[J].Gynecologic Oncology,2013,131(1):174-181.
[13]Lv Z,Hu M,Zhen J,et al.Rac1/PAK1 signaling promotes epithelial-mesenchymal transition of podocytes in vitro via triggering beta-catenin transcriptional activity under high glucose conditions[J].The International Journal of Biochemistry&Cell Biology,2013,45(2):255-264.
[14]Patel S,Takagi KI,Suzuki J,et al.RhoG TPase activation is a key step in renal epithelial mesenchymal transdifferentiation[J].J Am Soc Nephrol,2005,16(7):1977-1984.
[15]Liang Y,He L,Yuan H,et al.Association between RUNX3 promoter methylation and non-small cell lung cancer:a meta-analysis[J].Journal of Thoracic Disease,2014,6(6):694-705.
[16]Murata Y,Minami Y,Iwakawa R,et al.ECT2amplification and overexpression as a new prognostic biomarker for early-stage lung adenocarcinoma[J].Cancer Science,2014,105(4):490-497.