Characterization Of PTPN12 Knock-out HeLa Stable Cell Line

The human protein tyrosine phosphatase PTP-PEST is coded by the PTPN12 gene,and its abnormal expression or activity is closely related to various diseases.However,the research reports are still rather limited to the function of PTPPEST and the progression of PTP-PEST in tumors,particularly cervical cancer.The PX458-PTPN12-sg RNA1 and PX459-PTPN12-sg RNA2 recombinant vectors which were constructed in the previous stage of the research group were cotransfected to HeLa cells in this study.The stable HeLa cell line with the PTPN12 gene knockout was obtained after screening and identification.We observed the interaction between HeLa cells with PTPN12 gene knockout by the hanging drop experiments.We found that the deletion of PTP-PEST weakened the adhesion between cells and had a significant effect on cell proliferation compared with wildtype cells.The deletion of PTP-PEST significantly increased the number of HeLa cells in mitosis as shown in Giemsa staining experiments.We analyzed the cell cycle by flow cytometry and found that the number of cells in the S phase of the PTPPEST-deficient HeLa cell line was significantly increased,indicating that the knockout of the PTPN12 gene promoted cell proliferation in vitro.The migratory and invasive abilities of PTP-PEST-depleted HeLa cells were significantly enhanced in the cell scratch and Transwell cell invasion assays.PTP-PEST had no significant effect on the clone formation ability of HeLa cells as shown in the soft agar clone formation experiments.The mice tumor-bearing experiments also showed that the tumorigenic ability of HeLa cells after PTPN12 gene knockout was not significantly improved in vivo.Notably,the spleens of mice inoculated with PTPN12 knockout cells were significantly enlarged compared to those of mice inoculated with wild-type HeLa cells.Though there were no significant differences in the tumor tissue after HE staining,the number of multinucleated giant cells in the spleen of mice inoculated with PTPN12 gene knockout cells increased significantly.The result prompts that HeLa cells lacking PTP-PEST may aggravate the inflammatory response of mice.Finally,we found that the PTP-PEST is closely related to the p ERK,FAK,and Ecadherin signaling pathways,suggesting that the PTP-PEST may affect the process of cell adhesion and cell migration.In the later stage,further research is required into the migration capacity of tumors in vivo.In summary,we successfully constructed a PTPN12 knockout HeLa stable cell line and characterized the cell line.The results showed that the deletion of PTP-PEST improved the proliferation,migration,and invasion ability of HeLa cells in vitro.There was no significant effect on the ability of clone formation and tumorigenic in vivo.This study preliminarily revealed the effect of PTP-PEST in HeLa cells and provided a valuable cell model for studying the biological function of PTP-PEST in tumor cells.