Association of Methylation of the RAR-β Gene with Cigarette Smoking in Non-Small Cell Lung Cancer with Southern-central Chinese Population

  • Li, Wen (Key Laboratory of Green Packaging and Application of Biological Nanotechnology of Hunan Province) ;
  • Deng, Jing (College of Packaging and Material Engineering, Hunan University of Technology) ;
  • Wang, Shuang-Shuang (Key Laboratory of Green Packaging and Application of Biological Nanotechnology of Hunan Province) ;
  • Ma, Liang (Key Laboratory of Green Packaging and Application of Biological Nanotechnology of Hunan Province) ;
  • Pei, Jiang (Key Laboratory of Green Packaging and Application of Biological Nanotechnology of Hunan Province) ;
  • Zeng, Xiao-Xi (Key Laboratory of Green Packaging and Application of Biological Nanotechnology of Hunan Province) ;
  • Tang, Jian-Xin (Key Laboratory of Green Packaging and Application of Biological Nanotechnology of Hunan Province)
  • Published : 2015.01.22


Pathogenesis of lung cancer is a complicated biological process including multiple genetic and epigenetic changes. Since cigarette smoking is confirmed as the most main risk factor of non-small cell lung cancer (NSCLC), the aim of this study was to determine whether tobacco exposure plays a role in gene methylation. Methylation of the RAR-${\beta}$ gene were detected using methylation-specific polymerase chain reaction in DNA from 167 newly diagnosed cases with NSCLC and corresponding 105 controls. A significant statistical association was found in the detection rate of the promoter methylation of RAR-${\beta}$ gene between NSCLC and controls ($x^2$=166.01; p<0.01), and hypermethylation of the RAR-${\beta}$ gene was significantly associated with smoking status (p=0.038, p<0.05). No relationship was found between RAR-${\beta}$ gene methylation and pathologic staging including clinical stage, cell type, gender and drinking (p>0.05), and the methylation of RAR-${\beta}$ gene rate of NSCLC was slightly higher in stages III+IV (80.0%) than in I+II (70.8%). Similar results were obtained for methylation of the RAR-${\beta}$ gene between squamous cell carcinoma (77.9%) and other cell type lung cancer (73.9%). These results showed that the frequency of methylation increased gradually with the development of clinical stage in smoking-associated lung cancer patients, and tobacco smoke may be play a potential role in RAR-${\beta}$ gene methylation in the early pathogenesis and process in lung cancer, particularly squamous cell carcinoma. Aberrant promoter methylation is considered to be a promising marker of previous carcinogen exposure and cancer risk.


Supported by : National Natural Science Foundation of China


  1. Maruyama R, Toyooka S, Toyooka KO, et al (2002). Aberrant promoter methylation profile of prostate cancers and its relationship to clinicopathological features. Clin Cancer Res, 8, 514-9.
  2. Shaw RJ, Liloglou T, Rogers SN, et al (2006). Promoter methylation of P16, ARbeta, E-cadherin, cyclin A1 and cytoglobin in oral cancer: quantitative evaluation using pyrosequencing. BrJ Cancer, 94, 561-8.
  3. Losi-Guembarovski R, Kuasne H, Guembarovski AL, et al (2007). DNA methylation patterns of the CDH1, RARB, and SFN genes in choroid plexus tumors. Cancer Genet Cytogenet, 179, 140-5.
  4. Jin M, Kawakami K, Fukui Y, et al (2009). Different histological types of non-small cell lung cancer have distinct folate and DN Amethylation levels. Cancer Sci. 100, 2325-30.
  5. Suzuki M, Yoshino I (2010). Aberrant methylation in non-small cell lung cancer. Surg Today. 40, 602-7.
  6. Missaoui N, Hmissa S, Dante R, Frappart L, et al (2010). Global DNA methylation in precancerous and cancerous lesions of the uterine cervix. Asian Pac J Cancer Prev, 11, 1741-4.
  7. Hawes SE, Stern JE, Feng Q, et al (2010). DNA hypermethylation of tumors from non-small cell lung cancer (NSCLC) patients is associated with gender and histologic type. Lung Cancer, 69, 172-9.
  8. Jin Y, Xu H, Zhang C, et al (2010). Combined effects of cigarette smoking, gene polymorp-hisms and methylations of tumor suppressor genes on non small cell lung cancer: a hospital-based case-control study in China. BMC Cancer, 10, 422-30.
  9. Chung JH, Lee HJ, Kim BH, et al (2011). DNA methylation profile during multistage progression of pulmonary adenocarcinomas. Virchows Arch, 459, 201-11.
  10. Zhang Y, Wang R, Song H, et al (2011). Methylation of multiple genes as a candidate biomarker in non-small cell lung cancer.Cancer Lett, 303, 21-8
  11. Chen C, Yin N, Yin B, Lu Q (2011). DNA methylation in thoracic neoplasms.Cancer Lett, 301, 7-16.
  12. Daniunaite K, Berezniakovas A, Jankevicius F, et al (2011). Frequent methylation of RASSF1 and RARB in urine sediments from patients with early stage prostate cancer. Medicina, 47, 147-53.
  13. Breitling LP, Yang R, Korn B, et al (2011). Tobacco-smoking-related differential DNA methylation: 27K discovery and replication. Am J Hum Genet, 88, 450-7.
  14. Leong KJ, Wei W, Tannahill LA, et al (2011). Methylation profiling of rectal cancer identifies novel markers of early-stage disease, Br J Surg, 98, 724-34
  15. Wang J, Zhao SL, Li Y, et al (2012). 4- (Methylnitrosamino)-1- (3-pyridyl)-1-butanone induces retinoic acid receptor $\beta$ hypermethylation through DNA methyltransferase 1 accumulation in esophageal squamousepithelial cells. Asian Pac J Cancer Prev, 13, 2207-12.
  16. Hassanein M, Callison JC, Callaway-Lane C, et al (2012). The state of molecular biomarkers for the early detection of lung cancer. Cancer Prev Res, 5, 992-1006.
  17. Lokk K, Vooder T, Kolde R, et al (2012). Methylation markers of early-stage non-small cell lung cancer. PLoS One, 7, 39813.
  18. Cheng Z, Wang W, Song YN, et al (2012). hOGG1, p53 genes, and smoking interactions are associated with the development of lung cancer. Asian Pac J Cancer Prev. 13, 1803-8.
  19. Gasche JA, Goel A (2012). Epigenetic mechanisms in oral carcinogenesis. Future Oncol, 8, 1407-25.
  20. Zhao X, Wang N, Zhang M, et al (2012). Detection of methylation of the RAR-$\beta$ gene in patients with non-small cell lung cancer. Oncol Lett, 3, 654-8.
  21. Scesnaite A, Jarmalaite S, Mutanen P, et al (2012). Similar DNA methylation Pattern in lung tumours from smokers and never-smokers with second-hand tobacco smoke exposure. Mutagenesis, 27, 423-9.
  22. Suzuki M, Shiraishi K, Eguchi A, et al (2013). Aberrant methylation of LINE-1, SLIT2, MAL and IGFBP7 in non-small cell lung cancer. Oncol Rep, 29, 1308-14.
  23. Zhao Y, Zhou H, Ma K, et al (2013). Abnormal methylation of seven genes and their associations with clinical characteristics in early stage non-small cell lung cancer. Oncol Lett, 5, 1211-8.
  24. Siegel R, Naishadham D, Jemal A (2013). Cancer statistics, 2013. CA Cancer J Clin, 63, 11-30.
  25. Zeilinger S, Kuhnel B, Klopp N, et al (2013) Baurecht H. Tobacco smoking leads to extensive genome-wide changes in DNA methylation. PLoS One, 8, 63812.
  26. Li W, Deng J, Tang JX (2014). Combined effects methylation of FHIT, RASSF1A and RAR$\beta$ genes on non-small cell lung cancer in the Chinese population. Asian Pac J Cancer Prev, 15, 5233-7.
  27. Wu XM, Chen Y, Shao Y, et al (2014). Association between cigarette smoking and RASSF1A gene promoter hypermethylation in lung cancer patients: a meta- analysis. Asian Pac J Cancer Prev, 15, 8451-4.
  28. Farkas SA, Vymetalkova V, Vodickova L, et al (2014). DNA methylation changes in genes frequently mutated in sporadic colorectal cancer and in the DNA repair and Wnt/$\beta$-catenin signaling pathway genes. Epigenomics, 6, 179-91.
  29. Walter K, Holcomb T, Januario T, et al (2014). Discovery and development of DNA methylation-based biomarkers for lung cancer. Epigenomics, 6, 59-72.

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