[An experimental study of recombinant adenovirus microsphere carrying antisense multidrug resistance-associated protein gene for reversing MRP of hepatocellular carcinoma in vitro].
OBJECTIVE
To conduct an in vitro study on the effect of recombinant adenovirus microsphere encapusulated antisense MRP (as-mrp) for use in the gene therapy to overcome drug resistance in hepatocellular carcinoma.


METHODS
Recombinant adenovirus microsphere encapusulated as-mrp was transfected into hepatocellular carcinoma multidrug resistance cells HepG2/ADM, the fluorescence intensity of transfected cells were observed at 48 hours and 120 hours after transfection. in vitro drug sensitivity was measured by MTT assay; the resistant index of andromycin resistant variants was determined by drawing the cell dosage reaction curves. The levels of MRP mRNA expression were detected by RT-PCR and the ratio of MRP mRNA/beta-actin was detected. Intracelluar rubidomycin (DNR) concertration was examined by flow cytometry (FCM).


RESULTS
More than 90% of the HepG2/ADM cells could be transfected when microspheres being 10 mg. Adv microsphere inhibited the expression of mRNA in HepG2/ADM and enhanced the sensitivity of HepG2/ADM to chemotherapeutic drug.


CONCLUSIONS
Recombinant adenovirus microsphere encapusulated as-mrp could effectively reverse HepG2/ADM cells, which would provide an experimental basis for the methods of reversing the multidrug resistance in human hepatocellular carcinoma.
Yu
Shao-hong
SH
Department of Hepatobiliary Surgery, Chongqing Fuling Center Hospital, Fuling 400800, China.
Yan
Lü-nan
LN
Zhang
Yan
Y
Gou
Xing-hua
XH
Han
Lei
L
Chen
Yong-bing
YB
D000256:Adenoviridae / Q000235:genetics*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology; D008863:Microspheres; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D016372:RNA, Antisense / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D014162:Transfection
D027425:ATP-Binding Cassette, Sub-Family C Proteins; D016372:RNA, Antisense; D011994:Recombinant Proteins