Colorectal cancer (CRC), commonly known as bowel cancer is the third most common cause of cancer-related deaths in the western world. Colorectal cancer is one of the leading malignancies worldwide. CRC has been reported to show geographical variation in its incidence, even within areas of ethnic homogeneity. The usual treatment is surgery and subsequent chemotherapy and radiotherapy. Cancer development and progression is dictated by series of alterations in genes such as tumor suppressor genes, DNA repair genes, oncogenes and others. In colorectal carcinogenesis disturbances different from mutations called an epigenetic regulation are also taken into consideration.
The aim of this study was to study the promoter hypermethylation of CpG islands of p16 gene in colorectal cancer patients among the Kashmiri population. The study included 70 surgically obtained colorectal samples among which 50 were obtained from colorectal cancer patients and 20 were histopathologically normal colorectal samples. All the samples were histopathologically confirmed before further processing. DNA was extracted from all the samples and was modified using bisulphite modification kit. Methylation-specific polymerase (MSP) chain reaction was used for analysis of the promoter hypermethylation status of p16 gene. The genetic analysis of the cases and controls by MSP- PCR method, for checking the promoter hypermethylation of CpG islands of p16 gene revealed that unlike other high risk regions, Kashmiri population has a different promoter hypermethylation profile of p16 gene. 66% of the cases showed p16 promoter hypermethylation while as 34% of the cases were nonhypermethylated. The study also revealed that 20% of the normal cases also had promoter hypermethylation of p16 gene and 80% did not showed promoter hypermethylation of p16 gene. The association of promoter hypermethylation with colorectal cancer was evaluated by χ2 (Chi square) test with Odds ratio and was found to be significant. Among 29 male cases and 21 female cases, the association of promoter hypermethylation with colorectal cancer was evaluated using Fisher’s exact test and was found to be significant in both males and females. Occurrence of p16 promoter hypermethylation was found to be unequally distributed in males and females with more frequency in males than in females but the difference was not statistically significant. When the frequency of p16 promoter hypermethylation was compared with clinical staging of the disease, p16 promoter hypermethylation was found to be certainly higher in Stage III/IV (83.33%) compared to Stage I/ II (56.25%) but the difference was not statistically significant. Also, the degree of p16 promoter hypermethylation increased with the increasing severity of the lesion but the difference was not again statistically significant.
These results clearly suggest that p16 aberrant promoter hypermethylation in Kashmiri population contributes to the process of carcinogenesis in colorectal cancer and is reportedly one of the commonest epigenetic changes in the development of human CRC. It also demonstrates that hypermethylation of p16 gene can be designated as epigenetic biomarker for the screening, diagnosis and prognosis of colorectal cancer. The data gives a clue that p16 gene expression can be readily and fully restored and growth rate of cancer cells decreased by treatment of cancer cells with demethylating agents and DNA methylation inhibitors
