The Expression Of HSP90β In Human Gastric Cancer And MDR-type Gastric Cancer Cell Line And The Influence Of HSP90β Antisense MRNA To Drug-sensitivity Of Digestive Cancer Cell Line

AIM: 1.To evaluate the expression of HSP90 β in human gastric cancer tissue and drug-resistant gastric cancer cell line. 2. Constructing eukaryotic expression vector of HSP90 P antisense mRNA, transfecting tumor cells of the digestive system, to evaluate the responsiveness of the transfected tumor cells to chemotherapeutic drugs.METHODS: The expression of HSP90 P in human gastric cancer tissue and drug-resistant cells was studied by SABC immunohistochemical staining Eukaryotic expression vector of HSP90 P antisense mRNA was constructed by inserting into EcoR I (3 prime) and BamH I (5 prime )sites of pcDNA multiclonal sites a 1 .0kb HSP90 cDNA( which has a EcoR I 5 prime and BamH I 3 prime )from the primary plasmid phHSP90 and the reconstructed expression vector was transfected into human gastric cancer cell line SGC7901, MDR type gastric cancer cell line SGC7901/VCR hepatic cancer cell line HCC7402 and esophageal cancer cell line Ec109 by lipofectamine. Positive clones were selected by G418 and was named H-SGC7901, H-SC3C7901/VCR,H-HCC7402 and H-Ec109 respectively. Dot blot and RNase protection assay was used to analyze the expression of HSP90 P antisense mRNA Western blot and/or SABC immunohistochemical staining was used to assay the changes of HSP90 P ,PGP and TOPO II MTT was used to evaluate drug-sensitrvity of the transfected cells. ADR accumulation was analyzed by flow cytometry.