Effect Of KAI1 Gene Overexpression On The Proliferation,Invasion And Migrationof Non-small Celllung Cancer Cells

Objective:To investigate the effectofKAI1 gene overexpression on the biologicalfunctionofnon-smallcelllung cancer(NSCLC)cellsand the relatedmolecularmechanisms.Methods:KAI1geneoverexpressionplasmidgroupandemptyvectorco ntaining negative controlgroup were constructed respectively.Lpofecta min2000wasusedtotransientlytransfectnon-smallcelllungcancerc ellA549.Real-timequantitativePCR andWesternblottingwereusedto detecttheexpressionofKAI1 atthegeneandproteinlevelinthegro upofKAI1overexpressiongroup,emptyvectorgroupandblankgroup;CCK-8methodwasusedtodetecttheproliferationcapabilityofKAI1 g enebeforeandafteroverexpression.Theinvasionandmigrationcapabi lityofKAI1 gene before and afteroverexpression weredetected bythe methodofTranswellInvasionandMigrationexperiment;Real-timequanti tativePCR and Westernblotwereused to detecttheRNA and protein expressionlevelsofKAI1,SlugandE-cadherinineachgroup.Immunoh istochemistrywasusedtodetecttheexpressionofKAI1,SlugandE-ca dherininhumannon-smallcelllungcancertissues.Results:TheexpressionofKAI1mRNA inKAI1 overexpressiongroupwas significantlyhigherthanthatinemptyvectorgroupandblankgroupafter transient transfection,the difference was statistically significant(P<0.05),while there was no statistically significantdifferences in the expressionofKAI1betweentheemptyvectorgroupandtheblankgroup(P>0.05);TheexpressionofKAI1proteininKAI1overexpressiongroup was significantlyhigherthanthatinemptyvectorgroupandblankcontrolgroup(P<0.05).TheresultsofCCK-8showedthattheproliferativecapabilityof KAI1 overexpressiongroupwassignificantlyhigherthanthatoftheempty vectorgroup and blankgroup(P <0.05),while there wasno significant difference between the empty vector group and blank group(P>0.05);Transwellinvasion experimentshowed thatthe number of cells penetratingMatrigelinKAI1 overexpressiongroupwassignificantlyhigher thanthatinemptyvectorgroupandblankgroup(P<0.05),whiletherewas nosignificantdifferencebetweentheemptyvectorgroupandblankgroup(P>0.05);Transwellmigrationexperimentsshowedthatthecellsnumberof KAI1 overexpressiongrouppassingthroughthepolycarbonatemicroporous membranesignificantlyincreasedcomparedwiththeemptyvectorgroup and the blankgroup,the difference wasstatisticallysignificant(P<0.05),whiletherewasnosignificantdifferencebetweentheemptyvectorgroup and blankgroup(P> 0.05);Real-time quantitative PCR andWestern blot results showed thatSlug expression was decreased and E-cadherin expression wasincreased in KAI1 overexpression group compared with emptyvectorgroupandblankgroup.Theresultsofimmunohistochemistry showedthattheexpressionratesofKAI1andEcadherininnon-smallcelllungcancertissueswerelowerthanslug.Conclusions:OverexpressionofKAI1genecansignificantlydecreasedthe capabilityofproliferation,invasion and migration ofnon-smallcelllung cancercells;KAI1geneoverexpressioncandecreasedslugexpressionand promotedE-cadherinexpression;KAI1genemayinhibittheinvasionand migrationofnon-smallcelllungcancercellsbyregulatingtheexpressionof EMT.