Effect of miR-324-5p on drug resistance of pancreatic cancer cells through regulating KLF3/JAK2/STAT3 pathway
OBJECTIVE To explore the effect of miR-324-5p on drug resistance of pancreatic cancer(PC)cells,elucidate its potential molecular mechanism and provide theoretical and experimental rationales for seeking new therapeutic targets for PC.METHODS Plasma samples were collected from PC patients before and after chemotherapy.And PC cells were treated with dif-ferent concentrations of gemcitabine(Gem).Quantitative real-time polymerase chain reaction(qRT-PCR)was employed for detecting the expression of miR-324-5p in PC cells and plasma of PC patients.Cells were transfected with miR-324-5p mimics or inhibitors and treated with a certain concentration of Gem.Flow cytometry was utilized for detecting cell apoptosis and Western blot for assessing the expressions of apoptosis-related proteins and examining the effect of miR-324-5p on cell apoptosis.IC50 of Gem in cells was detected by CCK-8.And Western blot was utilized for detecting the expressions of drug resistance-related pro-teins and observing the effect of miR-324-5p on drug resistance.To further explore the molecular mechanism of miR-324-5p involved in drug resistance,Western blot was utilized for detecting the expression of JAK2/STAT3 signaling pathway after trans-fecting KLF3 overexpression plasmid.RESULTS miR-324-5p was raised in PC cells treated with Gem and plasma of PC patients on chemotherapy.And miR-324-5p suppressed cell apoptosis and apoptosis-related protein expression in PC cells treated with Gem.miR-324-5p boosted cell drug resistance to Gem and enhanced the expressions of drug resistance-related proteins.KLF3 overexpression not only partially reversed the effects of miR-324-5p on cell apoptosis and apoptosis-related protein expres-sion,but also partially reversed the effects of miR-324-5p on drug resistance and drug resistance-related protein expression in cells.Both effects were achieved through regulating the signaling pathway of JAK2/STAT3.CONCLUSION miR-324-5p is correlated with drug resistance of PC cells through activating JAK2/STAT3 signal via KLF3.
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