Effects of LINC00839 on proliferation,apoptosis,and migration of colorectal cancer cells by regulating the miR-17-5p/WEE1 axis
Objective To investigate the effects of long non-coding RNA LINC00839 on the proliferation,apoptosis,and migration of colorectal cancer(CRC)cells by regulating the microRNA-17-5p(miR-17-5p)/serine/threonine protein kinase 1(WEE1)axis.Methods The expression levels of LINC00839,miR-17-5p,and WEE1 mRNA in CRC cell lines were detected to screen the optimal cell line.LS513 cells were separated into Control group,LINC00839 knockdown negative control group(si-NC group),LINC00839 knockdown group(si-LINC00839 group),LINC00839 knockdown+miR-17-5p inhibition negative control group(si-LINC00839+NC inhibitor group),and LINC00839 knockdown+miR-17-5p inhibitor expression group(si-LINC00839+miR-17-5p inhibitor group).qRT-PCR was applied to detect the expression levels of LINC00839,miR-17-5p,and WEE1 mRNA in cells.Dual luciferase reporter gene experiment was applied to verify the targeting relationship between LINC00839,miR-17-5p,and WEE1.MTT was applied to detect cell viability.Edu was applied to detect cell proliferation.Scratch experiment was applied to detect the migration ability of cells.Flow cytometry was applied to detect cell apoptosis.Western blot was applied to detect protein levels of WEE1,PCNA,Bax,Bcl-2,caspase-3,E-cadherin,and N-cadherin.Results In the CRC cell line,LINC00839 and WEE1 mRNA were highly expressed,while miR-17-5p was low expressed.LS513 cells were selected for the experiment.Dual luciferase reporter gene experiment confirmed a targeted relationship between miR-17-5p and LINC00839,WEE1.Knocking down LINC00839 up-regulated miR-17-5p expression and down-regulated WEE1 expression.Knocking down LINC00839 greatly reduced LS513 cell viability,Edu positive rate,scratch healing rate,and expression of PCNA,Bcl-2,N-cadherin,and WEE1 proteins(P<0.05),while greatly increased the expression of Bax,caspase-3,and E-cadherin proteins(P<0.05).Inhibiting the expression of miR-17-5p was able to reverse the inhibitory effect of LINC00839 knockdown on the malignant biological behavior of LS513 cells(P<0.05).Conclusion LINC00839 is up-regulated in CRC cells,and knocking down LINC00839 may inhibit CRC cell proliferation and migration by regulating the miR-17-5p/WEE1 axis,promoting cell apoptosis.
Colorectal cancerLong non-coding RNA LINC00839MicroRNA-17-5p/serine/threonine protein kinase 1 axisProliferationApoptosisMigration
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