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1、抑制GM130表達(dá)對(duì)胃癌細(xì)胞增殖、侵襲及遷移能力的影響馮陽(yáng)陽(yáng)，陳婕，牟玲，蘇琳茜，易永芬（重慶醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院病理學(xué)教研室，分子醫(yī)學(xué)與腫瘤研究中心，重慶，400016）[摘要]目的分析GM130在不同分化人胃癌細(xì)胞系的差異表達(dá)，利用小干擾RNA技術(shù)來沉默GM130基因，研究其對(duì)胃癌細(xì)胞生物學(xué)行為的影響。方法體外培養(yǎng)三株不同分化程度胃癌細(xì)胞系（高分化MKN-28、中分化SGC-7901、低分化MKN-45）WesternBlot和RT-PCR篩選出高表達(dá)GM130的細(xì)胞株MKN-45、SGC-7901，
2、設(shè)計(jì)并化學(xué)合成、轉(zhuǎn)染、篩選出針對(duì)GM130的小干擾RNA片段。通過MTT、Transwell、Matrigel侵襲實(shí)驗(yàn)，觀測(cè)下調(diào)GM130后對(duì)胃癌細(xì)胞株的生物學(xué)行為影響，Westernblot檢測(cè)細(xì)胞中MMP-2、MMP-9蛋白的變化。結(jié)果WesternBlot和RT-PCR檢測(cè)GM130在MKN-45、SGC-7901細(xì)胞中呈現(xiàn)高表達(dá)。WesternBlot和qRT-PCR顯示在低分化胃癌MKN-45細(xì)胞中，GM130-siRNA-519轉(zhuǎn)染組GM130基因的表達(dá)水平顯著抑制（P﹤0.05）。與對(duì)照組
3、相比，抑制轉(zhuǎn)染組細(xì)胞其增殖、遷移能力明顯下降，穿膜細(xì)胞數(shù)明顯減少，MMP-2、MMP-9的蛋白表達(dá)水平顯著降低（P﹤0.05）。結(jié)論抑制GM130基因的表達(dá)下調(diào)可顯著降低胃癌細(xì)胞的增殖和體外侵襲轉(zhuǎn)移能力。[關(guān)鍵詞]GM130；RNA干擾；胃腫瘤；增殖；侵襲；遷移EffectsofGM130genesuppressiononproliferation,invasionandmigrationofhumangastriccarcinomacellFengYang-yang,ChenJie,MuLing,Su
4、Lin-qian,YiYong-fen(DepartmentofPathology，MolecularMedicineandCancerResearchCenter，ChongqingMedicalUniversity，Chongqing，400016，China)[Abstract]ObjectiveToexploretheexpressionofGM30inthedifferentiationofgastriccarcinomacelllinesandinvestigatetheeffectofsm
5、allinterferenceRNA-mediatedsilcencingoftheGM130geneoncellproliferation,invasionandmigrationofhumangastriccarcinomaMKN-45cellsinvitro.MethodsThreedifferentdifferentiatedgastriccarcinomacelllines(MKN-28,SGC-7901andMKN-45)wereculturedinvitro.WesternblotandR
6、T-PCRanalyseswerecarriedoutinMKN-45cellsandSGC-7901cells,whichhadhighGM130expressionlevels.ThedesignedandchemicallysynthesizedsiRNAtargetingtheGM130genewastransfectedintoMKN-45cells.TheeffectofGM130knockdownonMKN-45cellsproliferationwasdeterminedbyMTTass
7、ay,cellsmigrationwasanalyzedbyTranswellmigrationassay,theinvasionofcellswasdeterminedbyMatrigelinvasionassay.TheexpressionofMMP-2and-9wasalsodetectedattheproteinlevel.ResultsMKN-45andSGC-7901cellsshowedthehigherexpressionofGM130attheproteinandmRNAlevels,
8、soWesternblotwascarriedoutinMKN-45cells,whichhadinvasiveandmetastaticpotential.TheGM130-siRNA-519significantlydecreasedGM130proteinlevelcomparedtotwoothersequencesofthesiRNAtargetingGM130andscrambled.ThedistinctdecreaseofG
