| Objective: Heterogeneous nuclear ribonucleoprotein K(hnRNP K)is a member of the hnRNPs family,which is composed of a class of multifunctional protein family molecules that can regulate transcription,RNA selective splicing,translation,post-translation modification and cell signal transduction.In addition,a large number of studies have shown that hnRNP K is abnormally expressed in a variety of tumor cells and is closely related to the occurrence,development,stage and drug resistance of tumors.Our previous studies confirmed that hnRNP K is highly expressed in gastric cancer,and the high expression of hnRNP K is related to poor prognosis of patients,but the role of hnRNP K in gastric cancer is still unclear.In this study,the effect of hnRNP K on gastric cancer cells was studied from multiple aspects including proliferation,migration,invasion and drug resistance by knocking down the expression of hnRNP K protein in gastric cancer cells.Methods: In this study,the localization of hnRNP K in cells was detected by cellular immunofluorescence,and the expression of hnRNP K and related molecules in gastric cancer cells was detected by Western Blot.HnRNP Kinterference vector was constructed,and RNA interference method knocked down the expression of hnRNP K in human gastric cancer SGC-7901.The effects of hnRNP K on the proliferation,migration,invasion and drug resistance of gastric cancer cells were detected by MTT test,scratch test,Transwell migration test,Transwell invasion test.So as to study the role of hnRNP K in gastric cancer.Results:1.Cell immunofluorescence assay showed that hnRNP K was mainly located in the nucleus of SGC-7901 cells,with a small amount of expression in the cytoplasm.However,hnRNP K was only expressed in the nucleus of GES-1cells and almost not in the cytoplasm.2.Western Blot assay confirmed that hnRNP K shRNA 3 had the highest interference efficiency for hnRNP K.3.The transfection experiment showed that the number of cells in the hnRNP K interference group was significantly lower than that in the untreated group and the empty plasmid group,and the difference was statistically significant(p<0.05).The inhibitory effect was positively correlated with shRNA level,suggesting that hnRNP K could promote the proliferation of gastric cancer cells.4.Determined by MTT results showed: hnRNP K interference group compared with the empty plasmid group,untreated group,growth slowed significantly,no obvious increase in the number of living cells,proliferationability decreased significantly(P < 0.05),the difference was statistically significant,the empty plasmid group and untreated group,no significant difference,no statistically significant difference(P < 0.05),a further sign of hnRNP K can promote gastric cancer cell proliferation.5.Scratch test results showed that the migration distance of cells in the empty plasmid group and the untreated group was significantly greater than that in the hnRNP K interference group.The difference was statistically significant(p<0.05).There was no significant difference in cell migration distance between the empty plasmid group and the untreated group(p<0.05).6.Migration experiment results showed that: compared with the empty plasmid group and the untreated group,the number of cells migrated in the hnRNP K interference group was less,and the difference was statistically significant(p<0.05).There was no significant difference in the number of migrated cells between the empty plasmid group and the untreated group(p<0.05).7.Invasion experiment results showed that the number of invasion cells in the hnRNP K interference group was less than that in the empty plasmid group and the untreated group,and the difference was statistically significant(p<0.05).There was no significant difference in the number of invasion cells between the empty plasmid group and the untreated group(p<0.05).8.There was a certain difference in cell morphology between SGC-7901 and cisplatin resistant SGC-7901(SGC-7901/DDP).The cells ofSGC-7901/DDP were polygonal and had more abundant cytoplasm.SGC-7901 cells are polygonal and spindle-shaped with relatively few cytoplasm.9.Immunofluorescence experiments showed that hnRNP K was more expressed in the nucleus and cytoplasm of gastric cancer cell line SGC-7901/DDP,while hnRNP K was also expressed in the nucleus and cytoplasm of gastric cancer cell line SGC-7901,but the expression level was lower than that of SGC-7901/DDP cells,and the cytoplasm distribution was lower.10.Western Blot showed that the expression of hnRNP K in gastric mucosa GES-1 cells was lower than that in gastric cancer cells SGC-7901 and gastric cancer cells SGC-7901/DDP,and the difference was statistically significant(p<0.05).However,the expression of hnRNP K in gastric cancer cells SGC-7901/DDP was lower than that in gastric cancer cells SGC-7901/DDP,and the difference was statistically significant(p<0.05).Conclusion: 1.Knockdown of hnRNP K can inhibit the proliferation,migration and invasion of human gastric cancer cells SGC-7901.2.The increased expression of hnRNP K in SGC-7901/DDP cells and the increased cytoplasmic distribution suggested that hnRNP K was related to the drug resistance of gastric cancer cells SGC-7901. |
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