CDH13 promoter methylation regulates cisplatin resistance of non-small cell lung cancer cells

Wang,Yan; Zhang,Lei; Yang,Jiasheng; Li,Bin; Wang,Jun

doi:10.3892/ol.2018.9325

CDH13 promoter methylation regulates cisplatin resistance of non-small cell lung cancer cells

Authors:
- Yan Wang
- Lei Zhang
- Jiasheng Yang
- Bin Li
- Jun Wang
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Affiliations: Guangdong Second Provincial General Hospital, Guangzhou, Guangdong 510317, P.R. China, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510260, P.R. China
Published online on: August 20, 2018 https://doi.org/10.3892/ol.2018.9325
Pages: 5715-5722

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Abstract

Reversing cisplatin resistance of lung cancer cell line A549/DDP through recovering cadherin 13 (CDH13) expression by demethylation was investigated in the current study. RT-PCR was used to measure CDH13 expression in lung cancer A549 and A549/DDP cells with or without 5-Aza-CdR intervention. Methylation-specific PCR was used to detect CDH13 methylation. MTT assay and flow cytometry were used to measure the effects of cisplatin on inhibiting cell proliferation, apoptosis, and the reversal of cisplatin resistance. The IC50 value of cisplatin for A549 and A549/DDP cells was 3.278±0.532 and 28.341±1.435 µmol/l, respectively (P<0.05). The cisplatin-resistance index of A549/DDP cells was up to 8.65. After 2.5, 10, or 40 µmol/l 5-Aza-CdR treatment, the apoptotic rates of A549/DDP cells were 9.4±0.86, 18.1±1.42 and 42±2.01%, respectively, which were significantly different to those of the control group (P<0.05). Methylation-specific PCR detected both methylation (M) and unmethylation (U) bands at CDH13 promoter region before 5-Aza-CdR intervention while it only detected an unmethylation band after the treatment with a higher concentration of 5-Aza-CdR, which indicates the transformation to unmethylation state. When 10 µmol/l 5-Aza-CdR was added, the IC50 of cisplatin to A549/DDP cells was 8.472±0.415 µmol/l, and cisplatin resistance was reversed by 3.35‑fold. CDH13 methylation is related to the cisplatin resistance of A549/DDP cells. 5-Aza-CdR can inhibit CDH13 methylation and recover CDH13 expression. With the increase in 5-Aza-CdR concentration, the unmethylation state of CDH13 is enhanced, which can strengthen the function of cisplatin inhibiting proliferation and apoptosis in A549/DDP cells.

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1	Drilon A, Sugita H, Sima CS, Zauderer M, Rudin CM, Kris MG, Rusch VW and Azzoli CG: A prospective study of tumor suppressor gene methylation as a prognostic biomarker in surgically resected stage I to IIIA non-small-cell lung cancers. J Thorac Oncol. 9:1272–1277. 2014. View Article : Google Scholar : PubMed/NCBI
2	Morisaki H, Yamanaka I, Iwai N, Miyamoto Y, Kokubo Y, Okamura T, Okayama A and Morisaki T: CDH13 gene coding T-cadherin influences variations in plasma adiponectin levels in the Japanese population. Hum Mutat. 33:402–410. 2012. View Article : Google Scholar : PubMed/NCBI
3	Chung CM, Lin TH, Chen JW, Leu HB, Yang HC, Ho HY, Ting CT, Sheu SH, Tsai WC, Chen JH, et al: A genome-wide association study reveals a quantitative trait locus of adiponectin on CDH13 that predicts cardiometabolic outcomes. Diabetes. 60:2417–2423. 2011. View Article : Google Scholar : PubMed/NCBI
4	Hulpiau P and van Roy F: Molecular evolution of the cadherin superfamily. Int J Biochem Cell Biol. 41:349–369. 2009. View Article : Google Scholar : PubMed/NCBI
5	Dames SA, Bang E, Haüssinger D, Ahrens T, Engel J and Grzesiek S: Insights into the low adhesive capacity of human T-cadherin from the NMR structure of Its N-terminal extracellular domain. J Biol Chem. 283:23485–23495. 2008. View Article : Google Scholar : PubMed/NCBI
6	Ciatto C, Bahna F, Zampieri N, Van Steenhouse HC, Katsamba PS, Ahlsen G, Harrison OJ, Brasch J, Jin X, Posy S, et al: T-cadherin structures reveal a novel adhesive binding mechanism. Nat Struct Mol Biol. 17:339–347. 2010. View Article : Google Scholar : PubMed/NCBI
7	Liu T, Qian WJ, Gritsenko MA, Camp DG II, Monroe ME, Moore RJ and Smith RD: Human plasma N-glycoproteome analysis by immunoaffinity subtraction, hydrazide chemistry, and mass spectrometry. J Proteome Res. 4:2070–2080. 2005. View Article : Google Scholar : PubMed/NCBI
8	Chen R, Jiang X, Sun D, Han G, Wang F, Ye M, Wang L and Zou H: Glycoproteomics analysis of human liver tissue by combination of multiple enzyme digestion and hydrazide chemistry. J Proteome Res. 8:651–661. 2009. View Article : Google Scholar : PubMed/NCBI
9	Philippova M, Joshi MB, Kyriakakis E, Pfaff D, Erne P and Resink TJ: A guide and guard: The many faces of T-cadherin. Cell Signal. 21:1035–1044. 2009. View Article : Google Scholar : PubMed/NCBI
10	Resink TJ, Philippova M, Joshi MB, Kyriakakis E and Erne P: Cadherins and cardiovascular disease. Swiss Med Wkly. 139:122–134. 2009.PubMed/NCBI
11	Andreeva AV, Kutuzov MA, Tkachuk VA and Voyno-Yasenetskaya TA: T-cadherin is located in the nucleus and centrosomes in endothelial cells. Am J Physiol Cell Physiol. 297:C1168–C1177. 2009. View Article : Google Scholar : PubMed/NCBI
12	Emond MR and Jontes JD: Inhibition of protocadherin-alpha function results in neuronal death in the developing zebrafish. Dev Biol. 321:175–187. 2008. View Article : Google Scholar : PubMed/NCBI
13	Haas IG, Frank M, Véron N and Kemler R: Presenilin-dependent processing and nuclear function of gamma-protocadherins. J Biol Chem. 280:9313–9319. 2005. View Article : Google Scholar : PubMed/NCBI
14	Magg T, Schreiner D, Solis GP, Bade EG and Hofer HW: Processing of the human protocadherin Fat1 and translocation of its cytoplasmic domain to the nucleus. Exp Cell Res. 307:100–108. 2005. View Article : Google Scholar : PubMed/NCBI
15	Hou R, Liu L, Anees S, Hiroyasu S and Sibinga NE: The Fat1 cadherin integrates vascular smooth muscle cell growth and migration signals. J Cell Biol. 173:417–429. 2006. View Article : Google Scholar : PubMed/NCBI
16	Ferber EC, Kajita M, Wadlow A, Tobiansky L, Niessen C, Ariga H, Daniel J and Fujita Y: A role for the cleaved cytoplasmic domain of E-cadherin in the nucleus. J Biol Chem. 283:12691–12700. 2008. View Article : Google Scholar : PubMed/NCBI
17	Shoval I, Ludwig A and Kalcheim C: Antagonistic roles of full-length N-cadherin and its soluble BMP cleavage product in neural crest delamination. Development. 134:491–501. 2007. View Article : Google Scholar : PubMed/NCBI
18	Mukherjee S, Tessema M and Wandinger-Ness A: Vesicular trafficking of tyrosine kinase receptors and associated proteins in the regulation of signaling and vascular function. Circ Res. 98:743–756. 2006. View Article : Google Scholar : PubMed/NCBI
19	Takeuchi T and Ohtsuki Y: Recent progress in T-cadherin (CDH13, H-cadherin) research. Histol Histopathol. 16:1287–1293. 2001.PubMed/NCBI
20	Sato M, Mori Y, Sakurada A, Fujimura S and Horii A: The H-cadherin (CDH13) gene is inactivated in human lung cancer. Hum Genet. 103:96–101. 1998. View Article : Google Scholar : PubMed/NCBI
21	Zhong YH, Peng H, Cheng HZ and Wang P: Quantitative assessment of the diagnostic role of CDH13 promoter methylation in lung cancer. Asian Pac J Cancer Prev. 16:1139–1143. 2015. View Article : Google Scholar : PubMed/NCBI
22	Kim H, Kwon YM, Kim JS, Lee H, Park JH, Shim YM, Han J, Park J and Kim DH: Tumor-specific methylation in bronchial lavage for the early detection of non-small-cell lung cancer. J Clin Oncol. 22:2363–2370. 2004. View Article : Google Scholar : PubMed/NCBI
23	Hanabata T, Tsukuda K, Toyooka S, Yano M, Aoe M, Nagahiro I, Sano Y, Date H and Shimizu N: DNA methylation of multiple genes and clinicopathological relationship of non-small cell lung cancers. Oncol Rep. 12:177–180. 2004.PubMed/NCBI
24	Toyooka KO, Toyooka S, Virmani AK, Sathyanarayana UG, Euhus DM, Gilcrease M, Minna JD and Gazdar AF: Loss of expression and aberrant methylation of the CDH13 (H-cadherin) gene in breast and lung carcinomas. Cancer Res. 61:4556–4560. 2001.PubMed/NCBI
25	Morisaki H, Yamanaka I, Iwai N, Miyamoto Y, Kokubo Y, Okamura T, Okayama A and Morisaki T: CDH13 gene coding T-cadherin influences variations in plasma adiponectin levels in the Japanese population. Hum Mutat. 33:402–410. 2012. View Article : Google Scholar : PubMed/NCBI
26	Chen F, Huang T, Ren Y, Wei J, Lou Z, Wang X, Fan X, Chen Y, Weng G and Yao X: Clinical significance of CDH13 promoter methylation as a biomarker for bladder cancer: A meta-analysis. BMC Urol. 16:522016. View Article : Google Scholar : PubMed/NCBI
27	Guo Q, Wang HB, Li YH, Li HF, Li TT, Zhang WX, Xiang SS and Sun ZQ: Correlations of promoter methylation in WIF-1, RASSF1A, and CDH13 genes with the risk and prognosis of esophageal cancer. Med Sci Monit. 22:2816–2824. 2016. View Article : Google Scholar : PubMed/NCBI
28	Grady WM, Willis J, Guilford PJ, Dunbier AK, Toro TT, Lynch H, Wiesner G, Ferguson K, Eng C, Park JG, et al: Methylation of the CDH1 promoter as the second genetic hit in hereditary diffuse gastric cancer. Nat Genet. 26:16–17. 2000. View Article : Google Scholar : PubMed/NCBI
29	Andreeva AV and Kutuzov MA: Cadherin 13 in cancer. Genes Chromosomes Cancer. 49:775–790. 2010.PubMed/NCBI
30	Lin YL, Xie PG and Ma JG: Aberrant methylation of CDH13 is a potential biomarker for predicting the recurrence and progression of non muscle invasive bladder cancer. Med Sci Monit. 20:1572–1577. 2014. View Article : Google Scholar : PubMed/NCBI
31	Widschwendter A, Ivarsson L, Blassnig A, Müller HM, Fiegl H, Wiedemair A, Müller-Holzner E, Goebel G, Marth C and Widschwendter M: CDH1 and CDH13 methylation in serum is an independent prognostic marker in cervical cancer patients. Int J Cancer. 109:163–166. 2004. View Article : Google Scholar : PubMed/NCBI
32	Yang J, Niu H, Huang Y and Yang K: A systematic analysis of the relationship of CDH13 promoter methylation and breast cancer risk and prognosis. PLoS One. 11:e01491852016. View Article : Google Scholar : PubMed/NCBI
33	Hibi K, Kodera Y, Ito K, Akiyama S and Nakao A: Methylation pattern of CDH13 gene in digestive tract cancers. Br J Cancer. 91:1139–1142. 2004. View Article : Google Scholar : PubMed/NCBI
34	Ren JZ and Huo JR: Correlation between T-cadherin gene expression and aberrant methylation of T-cadherin promoter in human colon carcinoma cells. Med Oncol. 29:915–918. 2012. View Article : Google Scholar : PubMed/NCBI
35	Toyooka S, Toyooka KO, Maruyama R, Virmani AK, Girard L, Miyajima K, Harada K, Ariyoshi Y, Takahashi T, Sugio K, et al: DNA methylation profiles of lung tumors. Mol Cancer Ther. 1:61–67. 2001.PubMed/NCBI
36	Birchmeier W and Behrens J: Cadherin expression in carcinomas: Role in the formation of cell junctions and the prevention of invasiveness. Biochim Biophys Acta. 1198:11–26. 1994.PubMed/NCBI
37	Gayet O, Loncle C, Duconseil P, Gilabert M, Lopez MB, Moutardier V, Turrini O, Calvo E, Ewald J, Giovannini M, et al: A subgroup of pancreatic adenocarcinoma is sensitive to the 5-aza-dC DNA methyltransferase inhibitor. Oncotarget. 6:746–754. 2015. View Article : Google Scholar : PubMed/NCBI
38	Yang AS, Doshi KD, Choi SW, Mason JB, Mannari RK, Gharybian V, Luna R, Rashid A, Shen L, Estecio MR, et al: DNA methylation changes after 5-aza-2′-deoxycytidine therapy in patients with leukemia. Cancer Res. 66:5495–5503. 2006. View Article : Google Scholar : PubMed/NCBI
39	Nguyen TT, Mohrbacher AF, Tsai YC, Groffen J, Heisterkamp N, Nichols PW, Yu MC, Lübbert M and Jones PA: Quantitative measure of c-abl and p15 methylation in chronic myelogenous leukemia: Biological implications. Blood. 95:2990–2992. 2000.PubMed/NCBI
40	Blum W, Schwind S, Tarighat SS, Geyer S, Eisfeld AK, Whitman S, Walker A, Klisovic R, Byrd JC, Santhanam R, et al: Clinical and pharmacodynamic activity of bortezomib and decitabine in acute myeloid leukemia. Blood. 119:6025–6031. 2012. View Article : Google Scholar : PubMed/NCBI
41	Cowan LA, Talwar S and Yang AS: Will DNA methylation inhibitors work in solid tumors? A review of the clinical experience with azacitidine and decitabine in solid tumors. Epigenomics. 2:71–86. 2010. View Article : Google Scholar : PubMed/NCBI
42	Nicholson LJ, Smith PR, Hiller L, Szlosarek PW, Kimberley C, Sehouli J, Koensgen D, Mustea A, Schmid P and Crook T: Epigenetic silencing of argininosuccinate synthetase confers resistance to platinum-induced cell death but collateral sensitivity to arginine auxotrophy in ovarian cancer. Int J Cancer. 125:1454–1463. 2009. View Article : Google Scholar : PubMed/NCBI
43	Kuang P, Li XF, Li B, Wang YS, Li JY and Zhou CC: Comparative proteomic analysis of human lung adenocarcinoma A549 and A549/DDP cells. Tumor. 32:170–176. 2012.
44	Qin X, Yu S, Xu X, Shen B and Feng J: Comparative analysis of microRNA expression profiles between A549, A549/DDP and their respective exosomes. Oncotarget. 8:42125–42135. 2017. View Article : Google Scholar : PubMed/NCBI
45	Ye LY, Hu S, Xu HE, Xu RR, Kong H, Zeng XN, Xie WP and Wang H: The effect of tetrandrine combined with cisplatin on proliferation and apoptosis of A549/DDP cells and A549 cells. Cancer Cell Int. 17:402017. View Article : Google Scholar : PubMed/NCBI
46	Lin YL, He ZK, Li ZG and Guan TY: Downregulation of CDH13 expression promotes invasiveness of bladder transitional cell carcinoma. Urol Int. 90:225–232. 2013. View Article : Google Scholar : PubMed/NCBI
47	Kontic M, Stojsic J, Jovanovic D, Bunjevacki V, Ognjanovic S, Kuriger J, Puumala S and Nelson HH: Aberrant promoter methylation of CDH13 and MGMT genes is associated with clinicopathologic characteristics of primary non-small-cell lung carcinoma. Clin Lung Cancer. 13:297–303. 2012. View Article : Google Scholar : PubMed/NCBI
48	Zhai X and Li SJ: Methylation of RASSF1A and CDH13 genes in individualized chemotherapy for patients with non-small cell lung cancer. Asian Pac J Cancer Prev. 15:4925–4928. 2014. View Article : Google Scholar : PubMed/NCBI