Abstract:
:AIM:To construct Hsp90 antisense RNA eukaryotic expression vector, transfect it into SGC7901 and SGC7901/VCR of MDR-type human gastric cancer cell lines, HCC7402 of human hepatic cancer and Ec109 of human esophageal cancer cell lines, and to study the cell cycle distribution of the gene transected cells and their response to chemotherapeutic drugs.METHODS:A 1.03kb cDNA sequence of Hsp90beta was obtained from the primary plasmid phHSP90 by EcoR I and BamH I nuclease digestion and was cloned to the EcoR I and BamH I site of the pcDNA by T4DNA ligase and an antisense orientation of Hsp90beta expression vector was constructed. The constructs were transfected with lipofectamine and positive clones were selected with G418. The expression of RNA was determined with dot blotting and RNase protection assay, and the expression of Hsp90 protein determined with western blot. Cell cycle distribution of the transfectants was analyzed with flow cytometry, and the drug sensitivity of the transfectants to Adriamycin (ADR), vincrinstine (VCR), mitomycin (MMC) and cyclophosphamide (CTX) with MTT and intracellular drug concentration of the transfectants was determined with flow cytometry.RESULTS:In EcoR I and BamH I restriction analysis, the size and the direction of the cloned sequence of Hsp90beta remained what had been designed and the gene constructs were named pcDNA-Hsp90.AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones all expressed Hsp90 anti-sense RNA. The expression of Hsp90 was down-regulated in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones. Cell cycle distribution was changed differently. In AH-SGC7901/VCR and AH-Ec109 cells, G(1) phase cells were increased; S phase and G(2) phase cells were decreased as compared with their parental cell lines. In AH-SGC7901 cell, G(1)phase cells were decreased, G(2) phase cells increased and S phase cells were not changed, and in AH-HCC7402 cells G(1), S and G(2) phase cells remained unchanged as compared with their parental cell lines. The sensitivity of AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 to chemotherapeutic drugs, the sensitivity of AH-SGC7901/VCR to ADR, VCR, MMC and CTX the sensitivity of AH-HCC7402 to ADR and VCR, and the sensitivity of Ec109 to ADR, VCR and CTX all increased as compared with their parental cell lines. The mean fluorescence intensity of ADR in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 was also significantly elevated (P < 0.05).CONCLUSION: Down-regulation of Hsp90 could change cell cycle distribution and increase the drug sensitivity of tumor cells.
journal_name
World J Gastroenteroljournal_title
World journal of gastroenterologyauthors
Liu XL,Xiao B,Yu ZC,Guo JC,Zhao QC,Xu L,Shi YQ,Fan DMdoi
10.3748/wjg.v5.i3.199keywords:
subject
Has Abstractpub_date
1999-06-01 00:00:00pages
199-208issue
3eissn
1007-9327issn
2219-2840journal_volume
5pub_type
杂志文章abstract::The main treatment of patients with non-alcoholic fatty liver disease (NAFLD) is life style modification including weight reduction and dietary regimen. Majority of patients are safely treated with this management and pharmacologic interventions are not recommended. However, a subgroup of NAFLD patients with non-alcoh...
journal_title:World journal of gastroenterology
pub_type: 社论
doi:10.3748/wjg.v23.i42.7495
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journal_title:World journal of gastroenterology
pub_type: 临床试验,杂志文章,多中心研究
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doi:10.3748/wjg.v20.i42.15837
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abstract:AIM:To evaluate the clinical characteristics of nonvariceal upper gastrointestinal hemorrhage (NGIH) in patients with chronic kidney disease (CKD). METHODS:From 2003 to 2010, a total of 72 CKD patients (male n = 52, 72.2%; female n = 20, 27.8%) who had undergone endoscopic treatments for NGIH were retrospectively iden...
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journal_title:World journal of gastroenterology
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doi:10.3748/wjg.v19.i22.3520
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v8.i4.580
更新日期:2002-08-01 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v25.i22.2776
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journal_title:World journal of gastroenterology
pub_type: 临床试验,杂志文章
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更新日期:2006-09-28 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.15.2156
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pub_type: 杂志文章
doi:10.3748/wjg.v20.i39.14510
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v13.i47.6446
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i23.3582
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i6.899
更新日期:2005-02-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.14.632
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v17.i15.2007
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v19.i15.2362
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pub_type: 杂志文章
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pub_type: 杂志文章
doi:10.3748/wjg.v16.i3.345
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v12.i32.5234
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v22.i29.6673
更新日期:2016-08-07 00:00:00
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pub_type: 杂志文章,评审
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更新日期:2006-04-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v12.i13.2075
更新日期:2006-04-07 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v12.i17.2667
更新日期:2006-05-07 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i34.5266
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.15.3178
更新日期:2009-07-07 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.14.2343
更新日期:2008-04-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 历史文章,杂志文章,评审
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更新日期:2014-11-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v20.i15.4407
更新日期:2014-04-21 00:00:00