IntroductionThe purpose behind the experiments performed in the study, KLF4 hypermethylation promoter inactivates its tumor suppressor function in cervical carcinogenesis, is to investigate the mechanism by which the KLF4 gene it is silenced in cervical carcinomas. Cervical cancer is responsible for the deaths of 250,000 women every year. The development of therapies for cervical cancer has been limited due to a lack of genetic and epigenetic data on the cancer-causing mechanism. The KLF4 gene is a transcriptional regulator of cell growth and differentiation. It works as a tumor suppressor in cervical cancer, but is found to be inactivated in cervical cancer. Overexpression of KLF4 protein is known to inhibit cervical cancer cell growth and tumor formation by activating a cell cycle suppressor. Hypermethylation of the promoter CpG island can result in transcriptional silencing of many tumor suppressor genes. Two CpG regions, BSQ1 and BSQ3, were examined in this experiment. Experimental methods and results Cervical cancer tissues and normal cervical tissues were collected from 24 patients newly diagnosed with primary cervical cancer, in order to perform the experiments described in the article. Experiments were also performed on the following human cervical carcinoma cell lines: HeLa, SiHa, C33A, and CaSki, purchased from one company. The researchers extracted genomic DNA from the collected samples. The DNA was then modified with bisulfite and amplified by PCR. The PCR product was then examined by gel electrophoresis to ensure that a single band was obtained, and then sequenced using Invitrogen. Methylation-specific PCR of bisulfate-treated DNA was then performed. This was done to check the consistency of the… center of the paper… if they had tested multiple tumor cell lines, they may have come to drastically different conclusions. Furthermore, the experiment drew some drastic conclusions about the silencing mechanism of KFL4 and how 5-Aza treatment affects cancer cell lines. Perhaps other treatments would have the same or even better effects on tumor cell lines. To improve the conclusions of the work, the researchers should have repeated these experiments on a much larger sample. Testing people with different forms and stages of cervical cancer would have been helpful. Testing people of different backgrounds could also have an effect on the conclusions drawn from the experiments. Overall, I think the study did a decent job of researching the mechanism by which the KLF4 gene is silenced in cervical cancer, but some changes could have been made to improve the study.