| Pancreatic cancer(PC)is a highly malignant tumor of digestive system,which is considered to have the worst clinical prognosis among all human cancers.In recent years,pancreatic cancer has made great progress in precision surgery and combination therapy.However,because of its special anatomical location,occult disease,strong invasion,low radical surgery rate,easy drug resistance,high recurrence,metastasis and other characteristics,its incidence rate and mortality continue to rise in the global scale,which has become a global public health problem.Therefore,we should spare no effort to further study the molecular mechanism involved in the occurrence and development of pancreatic cancer,explore effective molecular biomarkers and potential therapeutic targets,so as to improve the early detection rate and therapeutic effect,and improve the prognosis of pancreatic cancer patients.Long chain non-coding RNA(Lnc RNAs)have a chain length of no less than 200 nucleotides,do not have the potential to code proteins,and are known to participate in almost all processes of cancer cell initiation,growth,metastasis and chemotherapy resistance as basic regulators.KCNQ1OT1,as a member of Lnc RNAs,has been identified for abnormal expression in various types of tumors.However,after reviewing the literature,we found that the research on its role in pancreatic cancer and its regulatory mechanism has not been reported yet.Therefore,this study starts with the abnormal expression of KCNQ1OT1,mi R-216 a,and STAT1 in human PC cells.Based on this,it further explores the interaction relationship between KCNQ1OT1/mi R-216a/STAT1 axis,and finally analyzes the impact of this competitive endogenous RNA(ce RNA)network on the biological behavior of PC and its possible molecular mechanisms.In this study,we selected human PC cell lines SW1990,Bx PC-3,PANC-1,As PC-1,and pancreatic ductal epithelial cell line HPDE as experimental and control groups,respectively.The levels of KCNQ1OT1,mi R-216 a,and STAT1 m RNA were detected using q RT PCR,and the protein expression of STAT1 was detected using Western blot.The results showed that in PC cells,KCNQ1OT1 and STAT1 m RNA/protein were highly expressed,while mi R-216 a was low expressed.We speculate that KCNQ1OT1 and mi R-216 a have binding sites by querying the bioinformatics website(Starbase),and believe that STAT1 m RNA is highly likely a downstream target gene of mi R-216 a.In order to verify the relationship between KCNQ1OT1,mi R-216 a and STAT1,we used double Luciferase experiments to confirm that mi R-216 a can bind to KCNQ1OT1 or STAT1.Through silencing/overexpression treatment,it was confirmed that KCNQ1OT1 competitively inhibits mi R-216 a activity,STAT1 m RNA is the target gene of mi R-216 a,and KCNQ1OT1 regulates the mi R-216a/STAT1 axis.To investigate the impact of this ce RNA network on the proliferation and invasion of human PC cells and its possible molecular mechanisms,we used MTT experiments and Transwell experiments to detect the proliferation and invasion of cells.The results showed that knocking down KCNQ1OT1 or overexpressing mi R-216 a could inhibit the proliferation and invasion ability of PC cells,while inhibiting mi R-216 a or overexpressing STAT1 m RNA could to some extent reverse the effect of si-KCNQ1OT1 on PC cells.Finally,to confirm that this mechanism of action can inhibit the growth of xenograft tumors in vivo,we chose to construct a xenograft tumor model and re apply q RT PCR and Western blot to detect RNA/protein in cancer tissue.The results showed that knocking down KCNQ1OT1 in vivo can regulate tumor growth by affecting the expression of mi R-216a/STAT1 axis.In conclusion,through this experimental study,knockdown of KCNQ1OT1 may inhibit the proliferation and invasion of pancreatic cancer cells by regulating the mi R-216a/STAT1 axis.This mechanism may provide a new direction for the early diagnosis of human pancreatic cancer,and may become a new strategy for the treatment of pancreatic cancer. |
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