Experimental Study Of Paclitaxel On Human Esophageal Carcinoma Cell Line Eca109 In Virto

Expeiimenlal study of pacli4axel on human esophagealcaidnoma cdl line EcalO9 in virtoAhstractObjeclives: (1) To examine whether paditaxel is effective in g~xiwd~i inhibition ofht~nan esophageal carcinoma cells (EcalO9). (2) To confirm the hypothesis that therapeutic effects of padlitaxel on EcalO9 cells were mediated by induction of apoptosis and cell cycle arrest (3) To elucidate the relationship of apoptosis and cell cycle arrest induced by paclitaxel. (4)Bcl-2, p53, p2lWAFLOPlpmteins were examined to stady the molecular mechanism of apoptosis induced by pacitaxel inEcalO9 cells.Methods: (1) Htinan Esophageal carcinoma cells, EcalO9, were maintained in RPMJ 1640 containing 100/0 fetal bovine sem and 1% penicilhin-streptomycin. Cellsweregix~as monolayer culUjresin 5% CO2 andpassagedatintervals of 3 days (2) Cytotoxicily was evaluated by the MIT assay in 96-well plates at different times after treatment ~vith peditaxel 0,10,20,40,80 nmolIL All of the cytotoxicity and cell viability assays were performed in Uipiate. Concentrations inhiing the ~ih 5004(1C50) were calculated alter padlitaxel treatment (3) Cell morphology was studied by light microscopy and transmission elec~n microscope. (4) Cell-DNA distnl,ution was performed widr flow cytometer. EcalO9 cells including adherent cells and nonadhetent cells were harvested Single color flrnescernt flow cytomefly (Becton Dickinson,USA). Histogramswere W with M II sotw. The tw of a for Cell twwi (AO tetw me1 tw ds was for orceuswithe un pe ed de ~. be--orevents in the An wi we ed (5) be -- orbo4 P53,P21so the by or all now pe (Q bow ~ as x i s for fiDm at bo 3 gh --. The~ nd was wt anu ds or mp.05 were ed statishallyguResulis: (1) In vibo, po could ta the pe Of halre Cells. Thefor of M Cells M in a timed mann fortalre ce& be xi0 orpe to bolre was 15 nmax at 4sh. (z) beho bo of -- wt bolre cens ed whpe were for The cell vothe ~ eq tw of~ anct ch or tw tw was am fu InOfPholop inbo1re cem ed wh pe. (3) rtw pe analyS for tape can rt the cell tw at the GN tw. bolre cells ed withhigh wt (l5, 35 n mop nd at GN fonger bofor ed whbe con M (5 nmot/L).W cem eti GN a tyPed mp pebebe GJG,mp be (4) The bo oftw BcL and P53 he notw bo nd and ed cens. un can be the ~of P2l wnal.bebo: (l) haax is efhave in pe the orfor ~cen be (ha1op lt optwpe is an ta dtug5for esophageal caiuinonia. (2) Padlltaxel can induce apoptosis and block cell cycle ofEcalO9 cells. (3) The cell cycle an~st at (iIM seems to iequiied for pacitaxelinducedapoptosis .(4)Bcl-2andp53 eannotbei~latedtoapoptosisinducedby paclitaxel in EcalO9 cells. p21 WAFIXJPI might participate in apoptosis induced by paclitaxel.