Expression of PIWI-interacting RNA-47851 in gastric adenocarcinoma and its influence on proliferation
- VernacularTitle:PIWI蛋白相互作用RNA-47851在胃腺癌中的表达及其对增殖的影响
- Author:
Jinli ZHU
1
;
Xinyue QIAO
;
Xuebing YAN
;
Chenghai WANG
Author Information
- Keywords: gastric adenocarcinoma; PIWI-interacting RNA-47851; cell proliferation; prolif-eration index; survival prognosis
- From: Journal of Clinical Medicine in Practice 2024;28(1):20-27,36
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the expression and clinical pathological significance of PIWI-interacting RNA-47851(piR-47851)in gastric adenocarcinoma and its influence on prolifera-tion.Methods The expression of piR-47851 was detected in 79 gastric adenocarcinoma tissues by real time fluorescence quantitative polymerase chain reaction(qRT-PCR),and the correlation of piR-47851 expression level and clinical features with survival and prognosis were analyzed.The effect of piR-47851 on proliferation activity of gastric cancer cells was observed by cell proliferation experi-ments.Informatics websites were used to predict the downstream target genes of piR-47851.The wild-type and mutant plasmids for the 3'untranslated region(UTR)of MAPK1 gene were established,and a dual luciferase reporting system was used to verify that pi R-47851 binded to the 3'UTR of MAPK1 gene,thereby inhibiting MAPK1 protein synthesis.The effect of overexpression/silencing of piR-47851 on MAPK1 expression level was examined through qRT-PCR experiment.Rescue experiment was used to confirm the direct regulatory effect of piR-47851 on MAPK1 and explore the effect of piR-47851/MAPK1 on the proliferation of gastric cancer cells.The effect of reduced piR-47851 expression on the volume and weight of transplanted tumors was observed in nude mice by xenograft tumor experi-ments;the effect of reduced piR-47851 expression on the cell proliferation activity of xenograft tumors was observed by Ki-67 staining.Results The qRT-PCR experiment result showed that the expression level of piR-47851 in 79 gastric adenocarcinoma tissues was significantly higher than that in normal gastric tissues adjacent to the cancer(P＜0.05).The expression level of piR-47851 was significantly correlated with tumor size,degree of differentiation,and survival prognosis(P＜0.05).Bioinformatics suggested that there was a complementary binding site between piR-47851 and MAPK1 gene 3'UTR.The dual luciferase reporter gene experiment showed that piR-47851 was able to significantly inhibit fluorescence enzyme activity in the MAPK1 wild-type transfection group,while this inhibitory effect was not observed in the MAPK1 mutant group.The CCK-8 proliferation activity experiment showed that overexpression of piR-47851 was able to significantly increase the proliferation activity of gastric cancer cells;after reducing piR-47851,the proliferation activity of cells decreased significantly(P＜0.05).Rescue experiment indicated that after changing the ex-pression of piR-47851 and(or)MAPK1 and then detecting the expression level of MAPK1 by qRT-PCR,the MAPK1 could be regulated by piR-47851.Overexpression of MAPK1 could functionally reduce the proliferation promoting effect of piR-47851 on gastric cancer cells,while reducing MAPK1 expression could further enhance the proliferation activity of gastric cancer cells.In xeno-graft tumor experiments,reducing the expression of piR-47851 significantly resulted in smaller tumor growth volume and weight(P＜0.05).The proliferation index Ki-67 staining showed that the num-ber of proliferative active cells in the reduced piR-47851 expression group was significantly lower than that in the control group(P＜0.05),which further validated the promotional effect of piR-47851 on cell proliferation.Conclusion The expression level of pi R-47851 is increased in gastric adenocarcinoma tissues,which is closely related with tumor size and survival prognosis.The piR-47851 downregulates MAPK1 protein expression by binding to the 3'UTR of MAPK1,thereby promo-ting the proliferation of gastric cancer cells.
