RT Journal Article
SR Electronic
T1 Reversal of Adriamycin Resistance of Hepatocellular Carcinoma by Targeting with Recombined Adenovirus Carring Antisense mdrl RNA
JF Chinese Journal of Clinical Oncology
JO Cancer Biol Med
FD China Anti-Cancer Association
SP 32
OP 36
VO 3
IS 1
A1 Xiong Ding
A1 Ying Mei
A1 Yujun Shi
A1 Jianping Gong
A1 Xuhong Li
A1 Yong Peng
A1 Yong Liu
A1 Chang’an Liu
YR 2006
UL http://www.cancerbiomed.org/content/3/1/32.abstract
AB OBJECTIVE Chemotherapy is an important therapy for hepatocellular carcinoma (HCC). However, it is not effective in many cases due to recurrence and metastasis even if the initial treatment produces a response. Multidrug resistance (MDR) is considered to be one of the considerable causes. The aim of this study was to reverse MDR of HepG2/ADM cells by blocking mdrl with an adenovirus vector carrying antisense mdrl in a tumor transplantated in athymic mice.METHODS pCMVMv E was removed from the pshuttle vector. A 0.3 kb AFP promoter was inserted into the pshuttle vector and pCMV changed into pAFP. The pAFP and asmdrl PCR products were doubly digested with Kpn and Apal, the digested products were ligated by T4 ligase, the asmdrl gene was inserted into pAFP and a newly plasmid pAFP-asmdr1 was constructed. Following digestion with Pl-Scel/I-Ceu I, pAFP-asmdr1 was ligated with Adeno-× genome DNA and amplified in E.coli XL1-Blue. The HEK293 cells were transfected and virus collected. The HepG2 MDR cells (HepG2/ADM) were induced by graded resistance to ADM and were inoculated into athymic mice. After adeno-asmdrl was injected, the expression of mdrl -mRNA and the volume of the transplantated tumor and its cells were observed.RESULTS Following injection with Adeno-asmdrl, the tumor volume in the ADM+Adeno-asmdr1 group did not increase. However the tumor volume in the PBS plus ADM group did significantly increase (P<0.05). in the tumor xenograft cells, mdrl mRNA in the xenografts was assessed by RT-PCR and was found to be reduced at 1 week and 4 weeks in the ADM+asmdr1 group, but it was stable in the ADM group. It was only 20% in the ADM+asmdr1 group compared to the ADM group at the 4th week (P<0.05). Evidence of apoptosis was observed in the tumor xenograft cells treated with Adeno-asmdrl, but there was rare or no apoptosis in the group treated with ADM and PBS.CONCLUSION Adenovirus carrying antisense mdrl RNA can partially reverse the MDR of HepG2/ADM cells and inhibit tumor growth by downregulating mdrl mRNA resulting in tumor cell apoptosis.