沉默LncRNA NORAD通过抑制TRIP13增加自噬从而抑制去势抵抗前列腺癌细胞的多西他赛耐药OA

Objective:To investigate the effects and molecular mechanisms of long non-coding RNA(LncRNA)DNA damage activated(NORAD)on docetaxel resistance in castration-resistant prostate cancer(CRPC)cells by regulating thyroid hormone receptor interactor 13(TRIP13)and the autophagy pathway.Methods:The sensitivity of CRPC cell line PC3 and its docetaxel-resistant strain PC3-DR to docetaxel was compared.PC3-DR cells were divided into 8 groups,including Control group,NORAD silenced group,NO-RAD silenced negative control group,TRIP13 silenced group,TRIP13 silenced negative control group,NO-RAD silenced+TRIP13 overexpression group,NORAD silenced+TRIP13 overexpression negative control group,and NORAD silenced+3-MA(autophagy inhibitor)group.EdU assay was used to detect prolifera-tion,flow cytometry was used to detect apoptosis,scratch assay and Transwell assay were used to analyze mi-gration and invasion.qRT-PCR was used to detect the expression of LncRNA NORAD.Western blot was used to detect the expression of TRIP13,Beclin1,LC3-II/I,and p62 proteins.Each group of PC3-DR cells was treated with docetaxel,and EdU assay and flow cytometry were used to detect proliferation and apoptosis,re-spectively,to analyze the changes in docetaxel sensitivity.Results:Compared with the PC3 group,the PC3-DR group showed significantly reduced inhibition rates of proliferation,migration,and invasion,and reduced apoptosis promotion rate after docetaxel treatment(P<0.05).Compared with the Control group and NORAD silenced negative control group,the NORAD silenced group showed significantly reduced cell proliferation,migration,and invasion(P<0.05),with significantly downregulated expression of LncRNA NORAD and TRIP13,significantly upregulated LC3-II/I ratio and Beclin1,and significantly downregulated p62(all P<0.05).Compared with the Control group or NORAD silenced negative control group,the TRIP13 silenced group showed significantly downregulated expression of TRIP13,significantly upregulated LC3-II/I ratio and Bec-lin1,and significantly downregulated p62(all P<0.05).Overexpression of TRIP13 or 3-MA treatment could partially reverse the effects of NORAD silencing(all P<0.05).Additionally,the proliferation inhibition rate and apoptosis promotion rate of the NORAD silenced group or TRIP13 silenced group were significantly in-creased compared with the Control group(all P<0.05),and overexpression of TRIP13 or 3-MA treatment could partially reverse the increased effects of NORAD silencing on proliferation inhibition rate and apoptosis promotion rate(all P<0.05).Conclusion:Silencing LncRNA NORAD promotes the sensitivity of PC3-DR cells to docetaxel by inhibiting TRIP13 expression and activating the autophagy pathway,thereby reversing do-cetaxel resistance in CRPC cells.This provides a theoretical basis for targeting the LncRNA NORAD/TRIP13/autophagy axis in the treatment of drug-resistant prostate cancer.