Preliminary Study On The Function Of New Gene C1orf63

ObjectiveC1orf63 is a new gene, expressing an unknown-function protein which is 33kD. So far, there is not any study reports about the function of C1orf63. One report said, IL-2 withdrawal in human T lymphoblast can induce some molecular signature of cell cycle exit, and C1orf63 is an up-regulated gene. Although its function is not clear, we can speculate that C1orf63 may have some relationship to cell cycle. As far as study on the function of a new gene, there is not any fixed model, but we have some basic strategies, such as the location and expression, purification of its protein, transfect, over-expression, RNA interference, knock - down, screening interaction proteins and so on. When study a new certain gene, we must carry some certain strategies on it. To explore the biological function of C1orf63, we carry out four experimental parts. Firstly, we investigate the expression of C1orf63 gene in liver cancer and precancerous tissues,and the mRNA and protein level of C1orf63 gene in several liver cancer cell lines and gastric cancer cell lines. Secondly, we detect the expression of C1orf63 in gastric cancer cell MKN28 after RNA interference. Thirdly, we study the effects on cell cycle, apoptosis, and proliferation activity of gastric cancer cell MKN28 after silencing C1orf63. Finally, we construct recombinant plasmid pEGFP-N1- C1orf63, which laid the foundation for the further research of C1orf63.MethodsImmunohistochemical staining was used to detect the C1orf63 gene expression and localization in the liver cancer tissue. Interference clips was carried into MKN28 cell lines by liposomes. The mRNA and protein level was detected by Real-Time PCR and Western blot separately. Flow cytometry was adopted to analyze cell cycle and apoptosis after silencing C1orf63 gene. Cell proliferation activity was detected by MTT.ResultsC1orf63 genes are cytoplasm expression in liver cancer tissues and precancerous tissues. Whether in mRNA level or protein level, the expression of C1orf63 gene was higher in MKN28 cell, while lower in MKN45 cell. After effective interference of C1orf63, the G1 phase was blocked and the cell proliferation activity gone down in MKN28 cells. Moreover, it comes up late apoptosis.ConclusionC1orf63 gene was cytoplasm expressed both in human liver cancer tissues and precancerous tissues. The expression of mRNA and protein levels was higher in MKN28 cells. Effective interference of C1orf63 caused the G1 phase blocking and decreased cell proliferation activity in MKN28 cells, especially 48h after transfect. And it conforms to our guess from the reference. Besides, effective interference of C1orf63 led to late apoptosis.