The Study Of The Difference Of Period1 And Period2 Between Gloma Cell Line And Normal Cell Line Cultured In Vitro

Life activity and habits of plants and animals in nature and human are all different during the day and night. The circadian rhythm about 24-hour is generated by the internal clock. The clock oscillation is a positive and negative feedback loop formed by a series of genes. Per1 and Per2 as the core circadian clock genes are essential in circadian rhythm generation and maintenance . Per1 and Per2 genes, mutation,deletion or abnormal expression can cause the biological circadian rhythm disorder or disappeared, followed by diseases or to promote the development of the disease, to promote premature aging or cancer. Through transgenic technology to increase Per1or Per2 gene in tumor cells, that lead Per1 or Per2 gene to overexpress in tumor cells can inhibit the growth of tumor cells. Intracranial malignant glioma is the most common central nervous system tumor. It is not easy to cut it completely. And radiotherapy and chemotherapy are side effects, and do not play well.Objective: Research has found that Per1 or Per2 gene mutation or deletion is closely related to breast cancer, lung cancer and malignant lymphoma genesis and development. To date,there is no study about the relevant association between glioma and the circadian clock genes.Our study is object to explore the relationship between the glioma and the circadian clock gene Per1, Per2.Method: PMA is used to induced Per1gene and Per2 gene to express in rhythm. The cells for different time points, using immunocytochemistry and RT-PCR, cultured glioma cells and normal cells expression gene expression differences. Immunocytochemistry and RT-PCR are used to test the expression of Per1 and Per2 in C6 cell and NIH3T3 cell.Results: The expression of Per1 and Per2 are detected in C6 cell and NIH3T3 cell. We used the Image pro-Plus 6.0 detect the immunocytochemistry images and analyzed the data using analysis of variance. We found that the expression of Per1 and Per2 in C6 cell are both different at the different time after treated by PMA.And we found that the expression of Per1 and Per2 in NIH3T3 cell are both no difference at the different time after treated by PMA.We also found that the expression level of Per1 and Per2 in C6 cell are both lower than in NIH3T3 cell. Semi-quantitative RT-PCR was used to detect mRNA of Per1 and Per2 in NIH3T3 cells and C6 cells.We measured OD values by gel imaging system and analyzed the data useing analysis of variance. We concluded the results that transcription of Per1 and Per2 are the same with translation of Per1 and Per2.Conclusion: The expression of Per1 and Per2 gene exist in C6 cells and NIH3T3 cells. The expression of Per1 and Per2 gene in NIH3T3 cells showed a circadian rhythm approximately. The expression of Per1 and Per2 in C6 cells had no significant rhythm and at a lower level.