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| 1 | Down-regulation of Hsp90 could change cell cycle distribution and increase drug sensitivity of tumor cells显示文摘:AIM To construct Hsp90 antisense RNAeukaryotic expression vector, transfect it intoSGC7901 and SGC7901/VCR of MDR-type humangastric cancer cell lines, HCC7402 of humanhepatic cancer and Eel09 of human esophagealcancer cell lines, and to study the cell cycledistribution of the gene transected cells andtheir response to chemotherapeutic drugs.METHODS A I .03kb cDNA sequence of Hsp90Pwas obtained from the primary plasmid phHsp90by EcoR 1 and BamH I nuclease digestion andwas cloned to the EcoR 1 and BamH 1 site ofthe pcDNA by T4DNA ligase and an antisenseorientation of Hsp900 expression vector wasconstructed. The constructs were transfectedwith lipofectamine and positive clones wereselected with G418. The expression of RNA wasdetermined with dot blotting and RNaseprotection assay, and the expression of Hsp90protein determined with Western blot. Cell cycledistribution of the transfectants was analyzedwith flow cytometry, and the drug sensitivity ofthe transfectants to adriamycin (ADR ),vincrinstine (VCR ), mitomycin (MMC ) andcyclophosphamide (CTX ) with MTT andintracellular drug concentration of thetransfectants was determined with flowcytometry.RESULTS In EcoR 1 and BamH I restrictionanalysis, the size and the direction of the clonedsequence of Hsp900 remained what had beendesigned and the gene constructs were namedpcDNA-Hsp90. AH^SGC7901, AH^SGC7901/ VCR,AH-HCC7402 and AH-Eel09 cell clones allexpressed Hsp90 anti--sense RNA. Theexpression of Hsp90 was down--regulated in AHSGC7901, AH--SGC7901/ VCR, AH-HCC7402 andAH--Eel09 cell clones. Cell cycle distribution waschanged differently. In AH-SGC7901/ VCR andAH-Ec109 cells, G, phase cells were increased; Sphase and G, phase cells were decreased ascompared with their parental cell lines. In AHSGC7901 cell, G, phase cells were decreased, Qphase cells increased and S phase cells were notchanged, and in AH-HCC7402 cells G,, S and qphase cells remained unchanged as comparedwith their parental cell lines. The sensitivity ofAH--SGC7901, AH--SGC7901/ VCR, AH-HCC7402 andAH-Ec109 to chemotherapeutic drugs, thesensitivity ot AH--SGC7901/ VCR to ADR, VCR,MMC and CTX the sensitivity of AH-HCC7402 toADR and VCR, and the sensitivity of Eel09 toADR, VCR and CTX all increased as comparedwith their parental cell lines. The meanfluorescence intensity of ADR in AH--SGC7901,AH-SGC7901/ VCR, AH--HCC7402 and AH-Ec109was also significantly elevated (P< 0. 05).CONCLUSION Down-regulation of HsP90 couldchange cell cycle distribution and increase thedrug sensitivity of tumor cells. | Liu XL Xiao B Yu ZC Guo JC Zhao QC Xu L Shi YQ Fan DM | 1999 | World Journal of Gastroenterology1999，5，3： | 21 |
| 2 | Investigation of recombinant Schistosoma japonicum paramyosin fragments for immunogenicity and vaccine efficacy in mice显示文摘 | Zhang DM Pan WQ Qian L Duke M Shen LH McManus DP | 2006 | 第二军医大学学报2006，27，6： | 6 |
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| 14 | Buried or unburied K-wires for lateral condyle elbow fractures显示文摘 | McGonagle L Elamin S Wright DM | 2012 | Ann R Coll Surg Engl2012，94，7： | 1 |
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| 16 | Measuring signal-to-noise ratios in MR imaging显示文摘 | Kaufman L Kramer DM Crooks LE | 1989 | Radiology1989，173，： | 1 |
| 17 | Selective blockade of the mGluR1 receptor reduces traumatic neuronal injury in vitro and improves outcome after trauma显示文摘 | Faden AI OLeary DM Fan L | 2001 | Exp Neurol2001，167，2： | 1 |
| 18 | IL-17 mRNA in sputum of asthmatic patients:linking T cell driven inflammation and granulocytic influx?显示文摘 | Bullens DM Truyen E Coteur L | 2006 | Respir Res2006，7，： | 1 |
| 19 | Statistics on cancer in China:cancer registration in 2002 显示文摘 | Yang L Parkin DM Whelan S | 2005 | Eur J Cancer Prey2005，14，4： | 1 |
| 20 | A Mycobacterium ulcerans toxin, mycolactone, causes apoptosis in guinea pig ulcers and tissue culture cells 显示文摘 | George KM Pascopella L Welty DM Small PL | 2000 | Infect Immun2000，68，2： | 1 |
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