Issue
Expression profiles of proto-oncogene TWIST1 and tumor metastasis suppressor gene LASS2 in bladder cancer
Corresponding Author(s) : Zeynep Yegin
Cellular and Molecular Biology,
Vol. 64 No. 11: Issue 11
Abstract
Transcription factor proto-oncogene TWIST1 and tumor metastasis suppressor gene LASS2 have been reported to be involved in various carcinomas but their expression profiles and prognostic significances in bladder cancer are largely unknown. We aimed to determine these genes' expression levels both at mRNA and protein level in bladder cancer. mRNA expression levels of TWIST1 and LASS2 genes were examined using real-time quantitative PCR (qPCR) in human bladder tumors and paired normal adjacent tissues obtained from 44 patients. Protein expression profiles of both genes were detected by immunohistochemical staining in formalin-fixed and paraffin-embedded tissues from the same patients. The expression profiles of TWIST1 mRNA in bladder tumors were significantly lower than the normal adjacent tissues and linked to both the stage and the grade. The expression profiles of LASS2 mRNA in bladder tumors were also significantly lower than the normal adjacent tissues reflecting the potential tumor suppressor profile of the gene, independently from stage or grade. By immunohistochemistry, TWIST1 and LASS2 positive expression rates were found as 14.3% (6/42) and 38.1% (16/42), respectively. As potential molecular markers for bladder carcinoma, both TWIST1 and LASS2 transcripts seem to play role during the tumorigenesis and development of bladder cancer. Lack of a functional link and/or weak inverse link between TWIST1 and LASS2 transcripts and immunohistochemical findings may reflect the potential associations of transcription regulation mechanisms and merit further investigations. To the best of our knowledge, this is the first report investigating the combined expression profile of TWIST1 and LASS2 in bladder cancer both at mRNA and protein level.
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- Ploeg M, Aben KK, Kiemeney LA. The present and future burden of urinary bladder cancer in the world. World J Urol 2009; 27, 3: 289-93.
- Enokida H and Nakagawa M. Epigenetics in bladder cancer. Int J Clin Oncol 2008; 13(4):298-307.
- Levy M and Futerman AH. Mammalian ceramide synthases. IUBMB Life 2010; 62, 5: 347-56.
- Becker I, Wang-Eckhardt L, Yaghootfam A, Gieselmann V, Eckhardt M. Differential expression of (dihydro)ceramide synthases in mouse brain: oligodendrocyte-specific expression of CerS2/Lass2. Histochem Cell Biol 2008; 129, 2: 233-41.
- Bauer R, Voelzmann A, Breiden B, Schepers U, Farwanah H, Hahn I, Eckardt F, Sandhoff K, Hoch M. Schlank, a member of the ceramide synthase family controls growth and body fat in Drosophila. EMBO J 2009; 28, 23: 3706-16.
- Wang H, Wang J, Zuo Y, Ding M, Yan R, Yang D, Ke C. Expression and prognostic significance of a new tumor metastasis suppressor gene LASS2 in human bladder carcinoma. Med Oncol 2012; 29, 3: 1921-7.
- Zhao Q, Wang H, Yang M, Yang D, Zuo Y, Wang J. Expression of a tumor-associated gene, LASS2, in the human bladder carcinoma cell lines BIU-87, T24, EJ and EJ-M3. Exp Ther Med 2013; 5, 3: 942-946.
- Pan H, Qin WX, Huo KK, Wan DF, Yu Y, Xu ZG, Hu QD, Gu KT, Zhou XM, Jiang HQ, Zhang PP, Huang Y, Li YY, Gu JR. Cloning, mapping, and characterization of a human homologue of the yeast longevity assurance gene LAG1. Genomics 2001; 77, 1-2: 58-64.
- Xu XY, Pei F, You JF. TMSG-1 and its roles in tumor biology. Chin J Cancer 2010; 29, 7: 697-702.
- Xu X, Liu B, Zou P, Zhang Y, You J, Pei F. Silencing of LASS2/TMSG1 enhances invasion and metastasis capacity of prostate cancer cell. J Cell Biochem 2014; 115(4):731-43.
- Gort EH, Suijkerbuijk KP, Roothaan SM, Raman V, Vooijs M, van der Wall E, van Diest PJ. Methylation of the TWIST1 promoter, TWIST1 mRNA levels, and immunohistochemical expression of TWIST1 in breast cancer. Cancer Epidemiol Biomarkers Prev 2008; 17, 12: 3325-3330.
- Miraoui H and Marie PJ. Pivotal role of Twist in skeletal biology and pathology. Gene 2010; 468, 1-2: 1-7.
- Maestro R, Dei Tos AP, Hamamori Y, Krasnokutsky S, Sartorelli V, Kedes L, Doglioni C, Beach DH, Hannon GJ. Twist is a potential oncogene that inhibits apoptosis. Genes Dev 1999; 13, 17: 2207-2217.
- Puisieux A, Valsesia-Wittmann S, Ansieau S. A twist for survival and cancer progression. BrJ Cancer 2006; 94, 1: 13-17.
- Wallerand H, Robert G, Pasticier G, Ravaud A, Ballanger P, Reiter RE, Ferrière JM. The epithelial-mesenchymal transition-inducing factor TWIST is an attractive target in advanced and/or metastatic bladder and prostate cancers. Urol Oncol 2010; 28, 5: 473-479.
- Shen CH, Wu JD, Jou YC, Cheng MC, Lin CT, Chen PC, Tseng YS, Shi CS, Chen SY, Chang DC, Lee YR. The correlation between TWIST, E-cadherin, and beta-catenin in human bladder cancer. J BUON 2011; 16(4):733-7.
- Li J and Zhou BP. Activation of β-catenin and Akt pathways by Twist are critical for the maintenance of EMT associated cancer stem cell-like characters. BMC Cancer 2011; 11: 49.
- Kwok WK, Ling MT, Lee TW, Lau TC, Zhou C, Zhang X, Chua CW, Chan KW, Chan FL, Glackin C, Wong YC, Wang X. Up-regulation of TWIST in prostate cancer and its implication as a therapeutic target. Cancer Res 2005; 65(12): 5153-62.
- Cheng GZ, Zhang W, Wang LH. Regulation of cancer cell survival, migration, and invasion by Twist: AKT2 comes to interplay. Cancer Res 2008; 68, 4: 957-960.
- Zhao N, Sun BC, Zhao XL, Liu ZY, Sun T, Qiu ZQ, Gu Q, Che N, Dong XY. Coexpression of Bcl-2 with epithelial-mesenchymal transition regulators is a prognostic indicator in hepatocellular carcinoma. Med Oncol 2012; 29, 4: 2780-92.
- Xu Xiao-yan, You Jiang-feng, Pei Fei, Zhang Bo. Silencing of tumor metastasis suppressor gene 1 promotes invasion of prostate cancer cell in vitro and its molecular mechanisms. Journal of Peking University (Health Sciences) 2011; 06.
- Tang X, Xing J, Li W, Wu Z, Zhang K, Zheng J. Expression of transcription factor Twist1 in bladder urothelial carcinoma and its clinical significance. J BUON 2013; 18, 1: 211-9.
- Livak KJ and Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25, 4: 402-8.
- Jouppila-Mättö A, Närkiö-Mäkelä M, Soini Y, Pukkila M, Sironen R, Tuhkanen H, Mannermaa A, Kosma VM. Twist and snai1 expression in pharyngeal squamous cell carcinoma stroma is related to cancer progression. BMC Cancer 2011; 11:350.
- Galván JA, Helbling M, Koelzer VH, Tschan MP, Berger MD, Hädrich M, Schnüriger B, Karamitopoulou E, Dawson H, Inderbitzin D, Lugli A, Zlobec I. TWIST1 and TWIST2 promoter methylation and protein expression in tumor stroma influence the epithelial-mesenchymal transition-like tumor budding phenotype in colorectal cancer. Oncotarget 2015; 6(2): 874–885.
- Song YH, Shiota M, Yokomizo A, Uchiumi T, Kiyoshima K, Kuroiwa K, Oda Y, Naito S. Twist1 and Y-box-binding protein-1 are potential prognostic factors in bladder cancer. Urol Oncol 2012; 1-7.
- Hotz B, Arndt M, Dullat S, Bhargava S, Buhr HJ, Hotz HG. Epithelial to mesenchymal transition: expression of the regulators snail, slug, and twist in pancreatic cancer. Clin Cancer Res 2007; 13(16):4769-76.
- Sung CO, Lee KW, Han S, Kim SH. Twist1 Is Up-Regulated in Gastric Cancer-Associated Fibroblasts with Poor Clinical Outcomes. The American Journal of Pathology 2011; 179 (4).
- Gao XH, Yang XQ, Wang BC, Liu SP, Wang FB. Overexpression of twist and matrix metalloproteinase-9 with metastasis and prognosis in gastric cancer. Asian Pac J Cancer Prev 2013; 14(9):5055-60.
- Kong YH, Syed Zanaruddin SN, Lau SH, Ramanathan A, Kallarakkal TG, Vincent-Chong VK, Wan Mustafa WM, Abraham MT, Abdul Rahman ZA, Zain RB, Cheong SC. Co-Expression of TWIST1 and ZEB2 in Oral Squamous Cell Carcinoma Is Associated with Poor Survival. PLoS One 2015; 10(7):e0134045.
- Nordfors K, Haapasalo J, Sallinen PK, Haapasalo H, Soini Y. Expression of claudins relates to tumour aggressivity, location and recurrence in ependymomas. Histol Histopathol 2013; 28(9):1137-46.
- Nagaishi M, Nobusawa S, Tanaka Y, Ikota H, Yokoo H, Nakazato Y. Slug, twist, and E-cadherin as immunohistochemical biomarkers in meningeal tumors. PLoS One 2012; 7(9):e46053.
- Yang JZ, Lian WG, Sun LX, Qi DW, Ding Y, Zhang XH. High nuclear expression of Twist1 in the skeletal extramedullary disease of myeloma patients predicts inferior survival. Pathol Res Pract 2016; 212(3):210-6.
- Wushou A, Hou J, Zhao YJ, Shao ZM. Twist-1 up-regulation in carcinoma correlates to poor survival. Int J Mol Sci 2014; 15(12):21621-30.
- Yegin Z, Gunes S, Buyukalpelli R. Hypermethylation of TWIST1 and NID2 in tumor tissues and voided urine in urinary bladder cancer patients. DNA Cell Biol 2013; 32(7):386-92.
- Tseng WC, Chuang CW, Yang MH, Pan CC, Tarng DC. Krüppel-like factor 4 is a novel prognostic predictor for urothelial carcinoma of bladder and it regulates TWIST1-mediated epithelial-mesenchymal transition. Urol Oncol 2016; 34(11):485.e15-485.e24.
- Fan S, Niu Y, Tan N, Wu Z, Wang Y, You H, Ke R, Song J, Shen Q, Wang W, Yao G, Shu H, Lin H, Yao M, Zhang Z, Gu J, Qin W. LASS2 enhances chemosensitivity of breast cancer by counteracting acidic tumor microenvironment through inhibiting activity of V-ATPase proton pump. Oncogene 2013; 32(13):1682-90.
- Ke RH, Wang Y, Mao Y, Zhang J, Xiong J. Decreased expression of LASS2 is associated with worse prognosis in meningiomas. J Neurooncol 2014; 118(2):369-76.
- Su J, Yu W, Gong M, You J, Liu J, Zheng J. Overexpression of a Novel Tumor Metastasis Suppressor Gene TMSG1/LASS2 Induces Apoptosis via a Caspase-dependent Mitochondrial Pathway. J Cell Biochem 2015; 116(7):1310-7.
- Gu D, Jin H, Jin G, Wang C, Wang N, Hu F, Luo Q, Chu W, Yao M, Qin W. The asialoglycoprotein receptor suppresses the metastasis of hepatocellular carcinoma via LASS2-mediated inhibition of V-ATPase activity. Cancer Lett 2016; 379(1):107-16.
- Ruan H, Wang T, Yang C, Jin G, Gu D, Deng X, Wang C, Qin W, Jin H. Co-expression of LASS2 and TGF-β1 predicts poor prognosis in hepatocellular carcinoma. Scientific Reports 2016; 6, Article number: 32421.
- Wang H, Zhang W, Zuo Y, Ding M, Ke C, Yan R, Zhan H, Liu J, Wang J. miR-9 promotes cell proliferation and inhibits apoptosis by targeting LASS2 in bladder cancer. Tumour Biol 2015; 36(12):9631-40.
- Liu J, Wang H, Wang Y, Li Z, Pan Y, Liu Q, Yang M, Wang J. Repression of the miR-93-enhanced sensitivity of bladder carcinoma to chemotherapy involves the regulation of LASS2. Onco Targets Ther 2016; 9:1813-22.
- Perez A, Loizaga A, Arceo R, Lacasa I, Rabade A, Zorroza K, Mosen-Ansorena D, Gonzalez E, Aransay AM, Falcon-Perez JM, Unda-Urzaiz M, Royo F. A Pilot Study on the Potential of RNA-Associated to Urinary Vesicles as a Suitable Non-Invasive Source for Diagnostic Purposes in Bladder Cancer. Cancers 2014; 6(1):179-92.
References
Ploeg M, Aben KK, Kiemeney LA. The present and future burden of urinary bladder cancer in the world. World J Urol 2009; 27, 3: 289-93.
Enokida H and Nakagawa M. Epigenetics in bladder cancer. Int J Clin Oncol 2008; 13(4):298-307.
Levy M and Futerman AH. Mammalian ceramide synthases. IUBMB Life 2010; 62, 5: 347-56.
Becker I, Wang-Eckhardt L, Yaghootfam A, Gieselmann V, Eckhardt M. Differential expression of (dihydro)ceramide synthases in mouse brain: oligodendrocyte-specific expression of CerS2/Lass2. Histochem Cell Biol 2008; 129, 2: 233-41.
Bauer R, Voelzmann A, Breiden B, Schepers U, Farwanah H, Hahn I, Eckardt F, Sandhoff K, Hoch M. Schlank, a member of the ceramide synthase family controls growth and body fat in Drosophila. EMBO J 2009; 28, 23: 3706-16.
Wang H, Wang J, Zuo Y, Ding M, Yan R, Yang D, Ke C. Expression and prognostic significance of a new tumor metastasis suppressor gene LASS2 in human bladder carcinoma. Med Oncol 2012; 29, 3: 1921-7.
Zhao Q, Wang H, Yang M, Yang D, Zuo Y, Wang J. Expression of a tumor-associated gene, LASS2, in the human bladder carcinoma cell lines BIU-87, T24, EJ and EJ-M3. Exp Ther Med 2013; 5, 3: 942-946.
Pan H, Qin WX, Huo KK, Wan DF, Yu Y, Xu ZG, Hu QD, Gu KT, Zhou XM, Jiang HQ, Zhang PP, Huang Y, Li YY, Gu JR. Cloning, mapping, and characterization of a human homologue of the yeast longevity assurance gene LAG1. Genomics 2001; 77, 1-2: 58-64.
Xu XY, Pei F, You JF. TMSG-1 and its roles in tumor biology. Chin J Cancer 2010; 29, 7: 697-702.
Xu X, Liu B, Zou P, Zhang Y, You J, Pei F. Silencing of LASS2/TMSG1 enhances invasion and metastasis capacity of prostate cancer cell. J Cell Biochem 2014; 115(4):731-43.
Gort EH, Suijkerbuijk KP, Roothaan SM, Raman V, Vooijs M, van der Wall E, van Diest PJ. Methylation of the TWIST1 promoter, TWIST1 mRNA levels, and immunohistochemical expression of TWIST1 in breast cancer. Cancer Epidemiol Biomarkers Prev 2008; 17, 12: 3325-3330.
Miraoui H and Marie PJ. Pivotal role of Twist in skeletal biology and pathology. Gene 2010; 468, 1-2: 1-7.
Maestro R, Dei Tos AP, Hamamori Y, Krasnokutsky S, Sartorelli V, Kedes L, Doglioni C, Beach DH, Hannon GJ. Twist is a potential oncogene that inhibits apoptosis. Genes Dev 1999; 13, 17: 2207-2217.
Puisieux A, Valsesia-Wittmann S, Ansieau S. A twist for survival and cancer progression. BrJ Cancer 2006; 94, 1: 13-17.
Wallerand H, Robert G, Pasticier G, Ravaud A, Ballanger P, Reiter RE, Ferrière JM. The epithelial-mesenchymal transition-inducing factor TWIST is an attractive target in advanced and/or metastatic bladder and prostate cancers. Urol Oncol 2010; 28, 5: 473-479.
Shen CH, Wu JD, Jou YC, Cheng MC, Lin CT, Chen PC, Tseng YS, Shi CS, Chen SY, Chang DC, Lee YR. The correlation between TWIST, E-cadherin, and beta-catenin in human bladder cancer. J BUON 2011; 16(4):733-7.
Li J and Zhou BP. Activation of β-catenin and Akt pathways by Twist are critical for the maintenance of EMT associated cancer stem cell-like characters. BMC Cancer 2011; 11: 49.
Kwok WK, Ling MT, Lee TW, Lau TC, Zhou C, Zhang X, Chua CW, Chan KW, Chan FL, Glackin C, Wong YC, Wang X. Up-regulation of TWIST in prostate cancer and its implication as a therapeutic target. Cancer Res 2005; 65(12): 5153-62.
Cheng GZ, Zhang W, Wang LH. Regulation of cancer cell survival, migration, and invasion by Twist: AKT2 comes to interplay. Cancer Res 2008; 68, 4: 957-960.
Zhao N, Sun BC, Zhao XL, Liu ZY, Sun T, Qiu ZQ, Gu Q, Che N, Dong XY. Coexpression of Bcl-2 with epithelial-mesenchymal transition regulators is a prognostic indicator in hepatocellular carcinoma. Med Oncol 2012; 29, 4: 2780-92.
Xu Xiao-yan, You Jiang-feng, Pei Fei, Zhang Bo. Silencing of tumor metastasis suppressor gene 1 promotes invasion of prostate cancer cell in vitro and its molecular mechanisms. Journal of Peking University (Health Sciences) 2011; 06.
Tang X, Xing J, Li W, Wu Z, Zhang K, Zheng J. Expression of transcription factor Twist1 in bladder urothelial carcinoma and its clinical significance. J BUON 2013; 18, 1: 211-9.
Livak KJ and Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25, 4: 402-8.
Jouppila-Mättö A, Närkiö-Mäkelä M, Soini Y, Pukkila M, Sironen R, Tuhkanen H, Mannermaa A, Kosma VM. Twist and snai1 expression in pharyngeal squamous cell carcinoma stroma is related to cancer progression. BMC Cancer 2011; 11:350.
Galván JA, Helbling M, Koelzer VH, Tschan MP, Berger MD, Hädrich M, Schnüriger B, Karamitopoulou E, Dawson H, Inderbitzin D, Lugli A, Zlobec I. TWIST1 and TWIST2 promoter methylation and protein expression in tumor stroma influence the epithelial-mesenchymal transition-like tumor budding phenotype in colorectal cancer. Oncotarget 2015; 6(2): 874–885.
Song YH, Shiota M, Yokomizo A, Uchiumi T, Kiyoshima K, Kuroiwa K, Oda Y, Naito S. Twist1 and Y-box-binding protein-1 are potential prognostic factors in bladder cancer. Urol Oncol 2012; 1-7.
Hotz B, Arndt M, Dullat S, Bhargava S, Buhr HJ, Hotz HG. Epithelial to mesenchymal transition: expression of the regulators snail, slug, and twist in pancreatic cancer. Clin Cancer Res 2007; 13(16):4769-76.
Sung CO, Lee KW, Han S, Kim SH. Twist1 Is Up-Regulated in Gastric Cancer-Associated Fibroblasts with Poor Clinical Outcomes. The American Journal of Pathology 2011; 179 (4).
Gao XH, Yang XQ, Wang BC, Liu SP, Wang FB. Overexpression of twist and matrix metalloproteinase-9 with metastasis and prognosis in gastric cancer. Asian Pac J Cancer Prev 2013; 14(9):5055-60.
Kong YH, Syed Zanaruddin SN, Lau SH, Ramanathan A, Kallarakkal TG, Vincent-Chong VK, Wan Mustafa WM, Abraham MT, Abdul Rahman ZA, Zain RB, Cheong SC. Co-Expression of TWIST1 and ZEB2 in Oral Squamous Cell Carcinoma Is Associated with Poor Survival. PLoS One 2015; 10(7):e0134045.
Nordfors K, Haapasalo J, Sallinen PK, Haapasalo H, Soini Y. Expression of claudins relates to tumour aggressivity, location and recurrence in ependymomas. Histol Histopathol 2013; 28(9):1137-46.
Nagaishi M, Nobusawa S, Tanaka Y, Ikota H, Yokoo H, Nakazato Y. Slug, twist, and E-cadherin as immunohistochemical biomarkers in meningeal tumors. PLoS One 2012; 7(9):e46053.
Yang JZ, Lian WG, Sun LX, Qi DW, Ding Y, Zhang XH. High nuclear expression of Twist1 in the skeletal extramedullary disease of myeloma patients predicts inferior survival. Pathol Res Pract 2016; 212(3):210-6.
Wushou A, Hou J, Zhao YJ, Shao ZM. Twist-1 up-regulation in carcinoma correlates to poor survival. Int J Mol Sci 2014; 15(12):21621-30.
Yegin Z, Gunes S, Buyukalpelli R. Hypermethylation of TWIST1 and NID2 in tumor tissues and voided urine in urinary bladder cancer patients. DNA Cell Biol 2013; 32(7):386-92.
Tseng WC, Chuang CW, Yang MH, Pan CC, Tarng DC. Krüppel-like factor 4 is a novel prognostic predictor for urothelial carcinoma of bladder and it regulates TWIST1-mediated epithelial-mesenchymal transition. Urol Oncol 2016; 34(11):485.e15-485.e24.
Fan S, Niu Y, Tan N, Wu Z, Wang Y, You H, Ke R, Song J, Shen Q, Wang W, Yao G, Shu H, Lin H, Yao M, Zhang Z, Gu J, Qin W. LASS2 enhances chemosensitivity of breast cancer by counteracting acidic tumor microenvironment through inhibiting activity of V-ATPase proton pump. Oncogene 2013; 32(13):1682-90.
Ke RH, Wang Y, Mao Y, Zhang J, Xiong J. Decreased expression of LASS2 is associated with worse prognosis in meningiomas. J Neurooncol 2014; 118(2):369-76.
Su J, Yu W, Gong M, You J, Liu J, Zheng J. Overexpression of a Novel Tumor Metastasis Suppressor Gene TMSG1/LASS2 Induces Apoptosis via a Caspase-dependent Mitochondrial Pathway. J Cell Biochem 2015; 116(7):1310-7.
Gu D, Jin H, Jin G, Wang C, Wang N, Hu F, Luo Q, Chu W, Yao M, Qin W. The asialoglycoprotein receptor suppresses the metastasis of hepatocellular carcinoma via LASS2-mediated inhibition of V-ATPase activity. Cancer Lett 2016; 379(1):107-16.
Ruan H, Wang T, Yang C, Jin G, Gu D, Deng X, Wang C, Qin W, Jin H. Co-expression of LASS2 and TGF-β1 predicts poor prognosis in hepatocellular carcinoma. Scientific Reports 2016; 6, Article number: 32421.
Wang H, Zhang W, Zuo Y, Ding M, Ke C, Yan R, Zhan H, Liu J, Wang J. miR-9 promotes cell proliferation and inhibits apoptosis by targeting LASS2 in bladder cancer. Tumour Biol 2015; 36(12):9631-40.
Liu J, Wang H, Wang Y, Li Z, Pan Y, Liu Q, Yang M, Wang J. Repression of the miR-93-enhanced sensitivity of bladder carcinoma to chemotherapy involves the regulation of LASS2. Onco Targets Ther 2016; 9:1813-22.
Perez A, Loizaga A, Arceo R, Lacasa I, Rabade A, Zorroza K, Mosen-Ansorena D, Gonzalez E, Aransay AM, Falcon-Perez JM, Unda-Urzaiz M, Royo F. A Pilot Study on the Potential of RNA-Associated to Urinary Vesicles as a Suitable Non-Invasive Source for Diagnostic Purposes in Bladder Cancer. Cancers 2014; 6(1):179-92.