Guangli Chen, Shusheng Gong, Juan Ding, Yingpeng Liu, Linghui Luo, Jianting Wang, Pei Chen, Kaisheng Yan
{"title":"[反义perlecan cDNA稳定表达对Hep-2细胞增殖的抑制]。","authors":"Guangli Chen, Shusheng Gong, Juan Ding, Yingpeng Liu, Linghui Luo, Jianting Wang, Pei Chen, Kaisheng Yan","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To study the suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA.</p><p><strong>Method: </strong>In this study, the plasmids of recombination eukaryotic expression vector perlecan anti-sense cDNA (pAP) were transfected into Hep-2 cells by using cationic liposome (lipofectamine 2000) and divided into three groups: non-transfected group, WT group; transfection with no load carrier, neo group; and transfection with the pAP plasmid, pAP group. Semi quantify RT-PCR, western blot assay and MTT assay were used to detected the expression of perlecan mRNA and protein in the three groups; the level of cell proliferation; and the responsivity of basic fibroblast growth factor (bFGF).</p><p><strong>Result: </strong>It was showed that the expression of perlecan mRNA and protein were significantly reduced in the pAP group compared with WT group and ph beta Apr-neol transfected group ( P < 0.01). In the presence of 1 microg/L of bFGF in low serum (0.1% FCS), the pAP transfected cells showed a reduced proliferation rate (MTT assay) while the wild type cells and ph beta Apr-neol transfected cells grew rapidly.</p><p><strong>Conclusion: </strong>The growth of Hep-2 cells could be inhibited significantly by perlecan anti-sense cDNA plasmids transfection.</p>","PeriodicalId":79680,"journal":{"name":"Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology","volume":"20 21","pages":"984-7"},"PeriodicalIF":0.0000,"publicationDate":"2006-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA].\",\"authors\":\"Guangli Chen, Shusheng Gong, Juan Ding, Yingpeng Liu, Linghui Luo, Jianting Wang, Pei Chen, Kaisheng Yan\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To study the suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA.</p><p><strong>Method: </strong>In this study, the plasmids of recombination eukaryotic expression vector perlecan anti-sense cDNA (pAP) were transfected into Hep-2 cells by using cationic liposome (lipofectamine 2000) and divided into three groups: non-transfected group, WT group; transfection with no load carrier, neo group; and transfection with the pAP plasmid, pAP group. Semi quantify RT-PCR, western blot assay and MTT assay were used to detected the expression of perlecan mRNA and protein in the three groups; the level of cell proliferation; and the responsivity of basic fibroblast growth factor (bFGF).</p><p><strong>Result: </strong>It was showed that the expression of perlecan mRNA and protein were significantly reduced in the pAP group compared with WT group and ph beta Apr-neol transfected group ( P < 0.01). In the presence of 1 microg/L of bFGF in low serum (0.1% FCS), the pAP transfected cells showed a reduced proliferation rate (MTT assay) while the wild type cells and ph beta Apr-neol transfected cells grew rapidly.</p><p><strong>Conclusion: </strong>The growth of Hep-2 cells could be inhibited significantly by perlecan anti-sense cDNA plasmids transfection.</p>\",\"PeriodicalId\":79680,\"journal\":{\"name\":\"Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology\",\"volume\":\"20 21\",\"pages\":\"984-7\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2006-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
[Suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA].
Objective: To study the suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA.
Method: In this study, the plasmids of recombination eukaryotic expression vector perlecan anti-sense cDNA (pAP) were transfected into Hep-2 cells by using cationic liposome (lipofectamine 2000) and divided into three groups: non-transfected group, WT group; transfection with no load carrier, neo group; and transfection with the pAP plasmid, pAP group. Semi quantify RT-PCR, western blot assay and MTT assay were used to detected the expression of perlecan mRNA and protein in the three groups; the level of cell proliferation; and the responsivity of basic fibroblast growth factor (bFGF).
Result: It was showed that the expression of perlecan mRNA and protein were significantly reduced in the pAP group compared with WT group and ph beta Apr-neol transfected group ( P < 0.01). In the presence of 1 microg/L of bFGF in low serum (0.1% FCS), the pAP transfected cells showed a reduced proliferation rate (MTT assay) while the wild type cells and ph beta Apr-neol transfected cells grew rapidly.
Conclusion: The growth of Hep-2 cells could be inhibited significantly by perlecan anti-sense cDNA plasmids transfection.
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