Antisense expression of PKC alpha improved sensitivity of SGC7901/VCR cells to doxorubicin

作者：Wu Da Long^*; Sui Feng Ying; Du Cheng; Zhang Cheng Wen; Hui Bin; Xu Shui Ling; Lu Huan Zhang; Song Guo Jie

来源：World Journal of Gastroenterology, 2009, 15(10): 1259-1263.

DOI：10.3748/wjg.15.1259

• Multi-drug resistance
• Protein kinase C alpha
• SGC7901
• Gastric cancer

AIM: To explore whether antisense blocking of protein kinase C alpha (PKC alpha) would reverse multi-drug resistance (MDR) in the vincristine (VCR)-resistant human gastric cancer cell line SGC7901/VCR.
METHODS: SGC7901/VCR cells expressing antisense PKCa, SGC7901/VCR/aPKC, were established by transfection with a recombinant plasmid reversely inserted with PKC alpha cDNA. Empty vector (PCI-neo)-transfected cell clones, SGC7901/VCR/neo, served as the control. Western blot method was used to detect PKC alpha content in SGC7901, SGC7901/VCR, SGC7901/VCR/neo and SGC790l/VCR/aPKC cells, using PKC alpha-specific antibody. The sensitivity of SGC7901, SGC7901/VCR, SGC7901/VCR/neo and SGC7901/VCR/aPKC cells to doxorubicin (DOX) in vitro was determined by MTT assay. The uptake of DOX in these cells was detected with fluorescence spectrophotometer.
RESULTS: Western blot analysis showed that the PKC alpha protein level was about 8.7-fold higher in SGC7901/VCR cells than that in SGC7901 cells, whereas the protein expression of PKC alpha was reduced by 78% in SGC7901/VCR/aPKC cells when compared with the SGC7901/VCR cells. SGC7901/VCR/aPKC cells had a 4.2-fold increase in DOX cytotoxicity, accompanied by a 1.7-fold increase of DOX accumulation in comparison with SGC7901/VCR cells.
CONCLUSION: PKC alpha positively regulates MDR in SGC7901 cells, and inhibition of PKC alpha can partially attenuate MDR in human gastric cancer cells.

• 出版日期2009-3-14
• 单位嘉兴学院; 中国人民解放军军事医学科学院