Expression And Mechanism Of Lonp2 In Human Head And Neck Cancer

Objective: To investigate the role of longp2 gene(peroxisome lon)in human head and neck cancer,we knocked out the protein encoded by longp2 gene and determined the effects of longp2 gene on proliferation,apoptosis and cell cycle of human head and neck cancer cells.Methods:1.The slow virus infection was caused by FaDu cells of hypopharyngeal squamous cell carcinoma.The experimental group was infected with sh RNA slow virus with candidate lonp2 gene,while the control group was infected with shctrl negative slow virus.The positive genes were screened by high content screening method: the effect of knockout of candidate genes(including lonp2)on the growth of FaDu cells and golim4 cells was detected by celigo experiment.The m RNA expression abundance of longp2 gene in FaDu,cal27 and CNE-2Z cells was detected by real-time quantitative PCR,and the m RNA expression after gene knockout was detected to determine the interference effect of target cells.The apoptosis number of shlonp2 gene was detected by annexinv APC single staining method,so as to detect the correlation between lonp2 gene and apoptosis.PI FACS was used to detect the changes of FaDu cell cycle after knockout of longp2 gene.The proliferation of FaDu cells was evaluated by the colony forming ability of infected cells on the cell culture plate.In order to determine the interference of knockout of longp2 gene on target cells,Westernblot was performed in 293 T cells.Results:To determine the knockout status of longp2 gene in human head and neck cancer,the expression abundance of longp2 gene in FaDu,cal27 and CNE-2Z cells was detected by PCR.The results showed that longp2 gene was highly expressed in FaDu,cal27 and CNE-2Z cells.Sh RNA lentivirus infection inhibited the expression of longp2 gene in FaDu cells(P < 0.05),and the transfection efficiency was 71.8%.5days after lentivirus infection,the expression of longp2 gene in FaDu cells was inhibited(P < 0.05),and the number of apoptotic FaDu cells increased significantly.The apoptotic rates of FaDu cells induced by annexin v-apc were 2.1%,2.3% and2.1% in shctrl group and 9.1%,9.6% and 9.8% in shlonp2 group,respectively.After 5days of culture,the proportion of G1,s,G2 phase cells in shctrl group was significantly higher than that in shctrl group,the number of S phase cells decreased(P< 0.05),the number of G1 phase cells increased(P < 0.05),and the number of G2 phase cells decreased(P < 0.05).The number of shlonp2 cells in the experimental group was significantly less than that in the control group(P < 0.05).WB test showed that the target could inhibit the expression of exogenous longp2 gene,which was an effective target.Conclusion: Lonp2 gene can inhibit the proliferation of tongue squamous cell carcinoma cell line FaDu,which is expected to be a therapeutic target for head and neck cancer.