| 【Objective】Through a large sample of prospective study to analyze the expression of PRDM14 and methylation status for the occurrence of lung cancer,to determine its clinical significance,to guide the treatment of individualized lung cancer【Methods】We prospectively collected 300 cases of stage T1-T2 non-small cell lung cancer tissue samples,combined with clinical and pathological data,expression and methylation of PRDM14 were detected by real-time quantitative PCR and pyrosequencing method,according to the analysis of clinical follow-up data after lung cancer occurrence and development of PRDM14 expression and methylation status of clinical significance for prognosis of lung cancer.At the cellular level to study the molecular mechanism of PRDM14 gene function and to participate in regulation.And to study the function of PRDM14 in animal level(in vivo)in lung cancer.【Results】1.The expression of PRDM14 in non-small cell lung cancer cell lines and tissues First,we first examined 40 lung cancer tumors and 40 adjacent nonmalignant lung tissue samples using PCR techniques to compare PRDM14 m RNA levels.We found that PRDM14 was mostly upregulated in lung cancer samples.We further compared the PRDM14 protein levels in lung cancer tissues and nonmalignant lung tissues by Western blot.The specificity of anti-PRDM14 antibody was confirmed by Western blot analysis of heterozygous expression of PRDM14 in 293 T cells.The results showed that the protein levels of PRDM14 in 40 cases of lung cancer were higher than those in non-malignant lung tissues with anti-PRDM14 antibody.The expression of PRDM14 in lung cancer cells was higher than that in non-malignant lung tissues,suggesting that PRDM14 may be involved in the occurrence and development of non-small cell lung cancer.2.Relationship between PRDM14 and lung cancer In order to verify the effect of PRDM14 on the prognosis of lung cancer,80 cases of cancerous and adjacent components were divided into PRDM14 overexpression group and PRDM14 low expression group by immunohistochemistry.The results showed that PRDM14 expression in lung cancer group was significantly higher than that in para-cancer group,32 cases of high expression of PRDM14 in lung cancer tissues and 11 cases of PRDM14 in adjacent normal control group,suggesting that PRDM14 expression was closely related to tumor metastasis The prognosis is positively correlated.3.Effects of interference and overexpression of PRDM14 on the proliferation of non-small cell lung cancer cell lines In order to study the expression of PRMD14 in non-small cell lung cancer,we first detected five non-small cell lung cancer cell lines(including H460,H358,H1975,H292,A549),and selected 1 × 105 cells The expression of PRDM14 was detected by western blot.We used three sh RNAs transfected with different sites of PRDM m RNA to transfected into NCI-H358,NCI-H1975 and A549 cells by lentivirus transfection technique.The protein level of PRDM14 was detected after 72 hours.The results showed that PRDM14 expression was significantly higher in non-small cell lung cancer cell lines than in normal bronchial epithelial cells.PRDM14 was silenced in NCI-H358 and NCI-H1975 cell lines.PRDM14 was overexpressed in A549 cells.Cell proliferation was observed after silencing,and cell proliferation was increased after overexpression.4.Effects of interference and overexpression of PRDM14 on apoptosis of non-small cell lung cancer cell lines We transfected si RNA and PRDM14 cells(PRDM14 si RNA3)or si RNA(sinc)by NCIH358 and NCI-H1975 cells;A549 cells infected with the virus or controlled the expression of lentiviral vectors(vectors).Cells were collected 24 hours after transfection with si RNA or lentivirus.PRDM14 was silenced in NCI-H358 and NCI-H1975 cell lines by interfering with PRDM14 in non small cell lung cancer cell lines,and PRDM14 was overexpressed in A549 cells The results showed that the interference of PRDM14 could promote the apoptosis of the cells,and the apoptosis of PRDM14 cells was decreased.5.Effects of interference and overexpression of PRDM14 on migration and invasion of non-small cell lung cancer cell lines In order to study the role of PRDM14 in the invasion of non-small cell lung cancer,we used Transwell assay to detect the difference of cell invasiveness.The results showed that the interference of PRDM14 reduced the migration of cells,and overexpression of PRDM14 promoted cell migration.6.Interference with PRDM14 inhibits in vivo tumorigenesis At the cellular level,interfering with PRDM14 significantly inhibited the proliferation of lung cancer cell beads,cloning formation,migration,and promoting tumor apoptosis.To further demonstrate the effect of PRDM14 on tumorigenesis in non-small cell lung cancer cells at the animal model level,we inoculated NCI-H358 cells and control groups on both sides of the back of nude mice.The results showed that the tumor volume of the shr DM14 group was significantly smaller than that of the control group.【Conclusion】PRDM14 plays an important role in the occurrence and development of non-small cell lung cancer,and PRDM14 promotes the proliferation,invasion and migration ability of non-small cell lung cancer.Reliable conclusions are obtained.The clinical significance for further non-small cell lung cancer targeted therapy provides a theoretical basis for further research,the relevant aspects of the drug PRDM14 inhibitor used in the treatment of lung cancer,guide the individual with excellent reference value. |
