| Purpose Epigenetic modification is closely related to the development of tumors,among which histone modification is a common way of epigenetic regulation of gene expression.Previous studies have shown that the PHF19 gene is involved in the process of histone modification,and is closely associated with the tumor development.This subject will research on the expression of PHF19 in nasopharyngeal carcinoma tissue,and the effect of PHF19 on the proliferation,migration and invasion of nasopharyngeal carcinoma cells,and to explore its role in the development of nasopharyngeal carcinoma.Methods 1.We used qRT-PCR to detect the relative expression level of PHF19 mRNA in 12 cases of nasopharyngeal carcinoma tissue and 12 cases of chronic inflammation of nasopharyngeal mucosa tissue;Immunohistochemistry was used to detect the relative expression level of PHF19 protein in 97 cases of nasopharyngeal carcinoma tissue and 58 cases of chronic inflammation of nasopharyngeal mucosa tissue.2.The lentiviral vectors containing PHF19 RNA interference fragment and empty fragment were designed and packaged.A human nasopharyngeal carcinoma cell line CNE1 was transfected with a lentiviral vector containing PHF19 RNA interference and empty fragment,respectively.They were used as PHF19-RNAi group and blank group,and nontransfected CNE1 as control group,a stable cell line was subsequently screened by puromycin.3.The relative expression level of PHF19 mRNA and protein in different groups were measured by qRT-PCR and western blot,respectively.4.The proliferation of the cells by MTT assay(represented by OD value)after cell culture for 24 h,48h,72 h and 96 h,separately.At the same time,the plate clone formation assay was carried out,and the number of clones was calculated when the clones were visible to the naked eye.5.The effects of PHF19 on migration of nasopharyngeal carcinoma cells were studied by cell wound scratch assay and transwell migration assay.The cell migration distance were measured and then relative mobility were calculated,and the number of penetrating cells were recorded.The relative expression level of E-cadherin mRNA and N-cadherin mRNA were detected in different groups of cells by qRT-PCR.6.The effect of PHF19 on the invasive ability of nasopharyngeal carcinoma cells was studied by transwell invasion assay,and the number of penetrating cells were recorded.Results 1.The relative expression level of PHF19 mRNA in nasopharyngeal carcinoma tissue and chronic inflammation of nasopharyngeal mucosa tissue were 0.621 ± 0.441 and 1.303 ± 0.868,respectively.Compared with chronic inflammation of nasopharyngeal mucosa tissue,the expression level of PHF19 mRNA was significant lower in nasopharyngeal carcinoma tissue(P < 0.05).The positive expression rate of PHF19 protein was 13.40% in 97 cases of nasopharyngeal carcinoma tissue and 81.03% in 58 cases of chronic inflammation of nasopharyngeal mucosa tissue.The positive expression rate of PHF19 protein in nasopharyngeal carcinoma was significant lower than that in the chronic inflammation of nasopharyngeal mucosa tissue(P < 0.05).2.The relative expression level of mRNA and protein of PHF19 in PHF19-RNAi group,blank group and control group were 0.173 ± 0.011,1.181 ± 0.195,1.00 ± 0.01 and 0.214 ± 0.007,0.512 ± 0.007,0.515 ± 0.009,respectively.Compared with the blank group and control group,the mRNA and protein expression level of PHF19 in the PHF19-RNAi group were significantly lower,and the difference was statistically significant(P < 0.05).3.The OD value of MTT were compared according to each group at different time points,which showed that no significant difference among different groups(P > 0.05).The number of cell clones in the PHF19-RNAi group,blank group and control group were 405 ± 20,416 ± 17 and 420 ± 21,respectively.There was no significant difference in the number of cell clones between each group,and the results were not statistically significant(P > 0.05).4.The relative mobility of the PHF19-RNAi group,blank group and control group at 24 hours were(76.25 ± 0.55)% ?(55.49 ± 0.84)%?(56.01 ± 0.45)%,respectively.The relative mobility of the cells in the PHF19-RNAi group was significant higher than that in the blank group and control group(P < 0.05).The transwell migration assay revealed that the numbers of penetrating cells in the PHF19-RNAi group,blank group and control group were 431 ± 14,324 ± 11 and 328 ± 10,respectively.The numbers of penetrating cells in the PHF19-RNAi group was significant higher than that in the blank group and control group(P < 0.05).5.The transwell invasion assay revealed that the numbers of penetrating cells in the PHF19-RNAi experimental group,blank group and control group were 120 ± 5?123 ± 7 and 125 ± 5,respectively.There was no significant difference in the number of penetrating cells between each group(P > 0.05).6.The relative expression level of Ecadherin mRNA and N-cadherin mRNA in the PHF19-RNAi group,blank group and control group were 0.493 ± 0.015,1.093 ± 0.099,1.00 ± 0.01 and 1.060 ± 0.079,1.137 ± 0.071,1.00 ± 0.01,respectively.The relative expression level of E-cadherin mRNA in the PHF19-RNAi group were significant lower than that in the blank group and control group(P < 0.05).While there was no significant difference in the relative expression level of N-cadherin mRNA among different groups(P > 0.05).Conclusion 1.The expression of PHF19 in nasopharyngeal carcinoma tissue was significantly lower than that in chronic inflammation of nasopharyngeal mucosa tissue.2.Stable PHF19 expression silencing could significantly promote the migration of nasopharyngeal carcinoma cells,but had no effect on the proliferation and invasion ability.3.PHF19 may be involved in the metastasis of nasopharyngeal carcinoma through the EMT pathway,and it is suggested that PHF19 may be a molecular marker associated with metastasis of nasopharyngeal carcinoma. |
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