Study On Action Mechanism Of ART Reversing Sotafenib Resistance In Human Hepatoma Cell Lines

OBJECTIVE:Aiming to establish a Human Hepatoma Bel-7402/Sora cell Line which resist Sorafenib and explore the Drug Resistant Mechanism.METHODS:Mtt assay was used to evaluate the cell inhibition of Bel-7402,we also calculate IC50.Sorafenib-resistant Human Hepatoma Cell(Bel-7402/Sora)was established by a high dose impact combined with continuous low dose concentration increasing method.Cell morphological was observed by an inverted microscope.Drug resistant multiple was also evaluated by Mtt assay.The expression of protein GRP78 and Beclinl was observed by Western-blot.We also evaluated the cell inhibition of Bel-7402 and Bel-7402/Sora with different dose of ART.The non-toxic concentration was further calculated.The reverse multiple and the relative reverse efficiency were also calculated.Different expression of protein GRP78 and Beclin1 under different concentration of Sora combine with toxic and non-toxic concentration ART was observed by western-blot.RESULTS:Bel-7402 and Bel-7402/Sora could monolayer grew and performed a scatteredSituation.Bel-7402 performed a ruled shape and evenly size.There was no bubble in the cytoplasm and it had a subtle shape.However,Bel-7402/Sora didn’t have a ruled shape and it had a varying size.Various bubbles and particles could observe the cytoplasm.The cell border seemed irregular and the cells gathered together.The growth rate of Bel-7402/Sora was also slower than Bel-7402.The IC50 was 37.7μmol/L(24h)and 8.133μmol/L(48h).There were no significant differences between 24h and 48h because of the cell traits and half-life of drugs.Thus 24h as serve as the criteria group.Comparing with Bel-7402,the resistance index of Bel-7402/Sora was 2.42.The expression of protein GRP78 and Beclinlrised as well.The IC50(24h)of Bel-7402/Sora which incubated under ART combined with Sora was 11.93μmol/L.The inverse index was 3.16 and the reverse efficacy rate was 116%.The expression of protein GRP78 and Beclin1 were also decreased.CONCLUSION:We successfully established the Human Hepatoma sorafenib resistant cell line Bel-7402/Sora.Protein GRP78 and Beclin1 were proven to be an accessory to formate the cell resistance of Sora,ART could decrease the expression of GRP78 and Beclinl to reverse the sorafenib resistance of Human Hepatoma carcinoma.