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体外转染KISS-1基因对食管鳞癌细胞EC-1侵袭及增殖能力的影响 被引量：4

Effect of KISS-1 on invasive potential and proliferation of esophageal squamous carcinoma cell line EC-1
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摘要 目的观察KISS-1基因对食管鳞癌细胞EC-1侵袭及增殖能力的影响，探讨KISS-1基因与食管鳞癌生物学行为的关系。方法检测KISS-1在EC-1、Eca109、EC9706和TE-1四种食管癌细胞株中的表达；利用脂质体介导在体外将KISS-1基因转染人食管鳞癌细胞EC-1，利用Western blot和RT-PCR方法检测转染之后KISS-1表达的改变，并采用Boyden小室体外侵袭实验及Mrrr、软琼脂克隆形成实验，观察KISS-1对食管鳞癌细胞侵袭及增殖能力的影响。结果4种食管癌细胞株的Western blot和RT-PCR检测结果显示：EC-1中KISS-1蛋白（0．715±0．109）及mRNA（0．670±0．176）表达均最低；转染之后Western blot和RT-PCR结果显示：KISS-1蛋白和mRNA在转基因组的表达分别为1．143±0．218和0．877±0．162，均显著高于转空质粒组（0．745±0．130，0．685±0．128；t=3．850，2．481，P均〈0．05）和对照组EC-1细胞（0．855±0．184，0．677±0．138；t=2．275，2．306，P均〈0．05）；Boyden小室检测细胞体外侵袭力实验发现转基因组在培养24、48和72h后的穿膜细胞数分别为91．8±11．7，117．8±11．1和139．2±11．8，均显著低于转空质粒组（118．1±14．7，141．7±13．2，162．2±22．7；t=3．153，4．215，3．569，P值均〈0．01）和对照组EC-1细胞（112．2±15．6，138．1±13．0，162．3±14．0；t=4．154，3．797，2．702，P值均〈0．05）。MTT检测结果显示，转基因组细胞在培养48h和72h后的增殖能力分别为0．517±0．127和0．394±0．137，与转空质粒组（0．636±0．186，0．513±0．150；t=2．054，2．709，P值均〈0．05）和对照组（0．646±0．135，0．511±0．153；f=2．276，2．205，P值均〈0．05）相比，细胞生长受到明显抑制；克隆形成实验结果显示，转基因组细胞的克隆形成数为157．2±36．4，明显低于转空质粒组（236．3±78．1；t=3．441，P〈0．01）和对照组（242．5±48．6；t=2．250，P〈0．05）。结论KISS-1基因在食管癌细胞株EC-1中可抑制细胞的体外侵袭能力及细胞的增殖能力。 Objective To investigate the effect of KISS-1 expression on the potential of invasion and proliferation of esophageal squamous carcinoma cell EC-1. Methods Protein and mRNA expressions of KISS-1 were evaluated by Western blot and RT-PCR in four esophageal carcinoma cell lines （EC-1, Eca109, EC9706 and TE-1）. Using liposome-mediated transfeetion, an eukaryotic expression vector （pcDNA3.1-KISS-1） of KISS-1 gene was transfeeted into EC-1 cells. Boyden chamber model, MTF and clone formation assay were used to detect the potential of invasion and proliferation. Results Western blot and RT-PCR showed a baseline low level of expression of KISS-1 protein （0.715±0. 109） and mRNA （0. 670±0. 176） in EC-1 cells, pcDNA3.1-KISS-1 expression vector was successfully transfected into EC-1 cells. Western blot and RT-PCR showed that the expression of KISS-1 protein （ 1. 143±0. 218） and mRNA （0. 877±0. 162 ） in EC-1 cells transfected with pcDNA3.1-KISS-1 were significantly higher than those transfected with the control vector pcDNA3. 1 （0.745 ± 0. 130,0. 685 ± 0. 128 ; t = 3. 850,2. 481, P 〈 0.05） and the control cells （0.855±0.184,0.677±0.138; t =2.275,2.306,P 〈0.05）. Boyden chamber analysis showed that the invasiveness of the cells transfected with KISS-1 at 24 h （91.8± 11.7 ） , 48 h （ 117.8± 11.1 ） and 72 h （ 139. 2 ±11.8） were significantly reduced than that of the cells transfected with the control vector peDNA3.1 （118.1±14. 7, 141.7±13.2, 162.2 ±22.7; t =3.153, 4. 215, 3.569, P〈0.01） and the control cells （112.2±15.6,138.1 ±13.0,162.3 ±14.0; t=4.154,3.797, 2. 702 ,P 〈 0. 05 ）. MTT showed that the proliferation potential of cells after transfection with KISS-1 at 48 h （0. 517±0. 127） and 72 h （0. 394 ±0. 137） were significantly reduced than that of cells transfected with the control vector pcDNA3.1 （ 0. 636 ± 0. 186, 0. 513 ± 0. 150 ; t = 2. 054, 2. 709, P 〈 0. 05 ） and the control cells （0.646 ±0. 135, 0.511 ±0. 153; t =2.276, 2.205, P 〈0.05）. Clone formation assay suggested that cells transfected with KISS-1 （157.2 ± 36. 4） showed significantly decreased clone formation than cells transfected with the control vector pcDNA3.1 （ 236. 3± 78.1 ; t = 3. 441, P 〈 0. 01 ） and the control cells （242. 5± 48. 6 ; t = 2. 250, P 〈 0. 05 ）. Conclusion KISS-1 gene inhibits the potential of invasion and proliferation of EC-1 cells.
作者 李娜 李珊珊 张红燕 轩小燕 郑献召 王丰 阎爱华
机构地区 郑州大学第一附属医院病理科河南省肿瘤病理重点实验室 新乡医学院病理教研室
出处 《中华病理学杂志》 CAS CSCD 北大核心 2009年第4期263-267,共5页 Chinese Journal of Pathology
基金 河南省科技攻关基金（072102310042）
关键词 食管肿瘤 转染 细胞系 肿瘤 KISS-1基因 Esophageal neoplasms Transfection Cell line, tumor KISS-1 gene
分类号 R686 [医药卫生—骨科学]
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参考文献9

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二级参考文献6

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中华病理学杂志
2009年 第4期

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