人MEG3基因质粒载体的构建及其对人胰腺癌SW1990细胞增殖的影响

doi:10.3760/cma.j.issn.1673-422X.2015.07.003

国际肿瘤学杂志 ›› 2015, Vol. 42 ›› Issue (7): 488-491.doi: 10.3760/cma.j.issn.1673-422X.2015.07.003

人MEG3基因质粒载体的构建及其对人胰腺癌SW1990细胞增殖的影响

陈瑞东,唐文,胡端敏

215004 苏州大学附属第二医院消化内科

出版日期:2015-07-08 发布日期:2015-05-26
通讯作者: 胡端敏 E-mail:huduanmin@163.com
基金资助:
苏州市科技计划（SYS201344）

Construction and inhibitory effect of MEG3 expression plasmid vector on the proliferation of human pancreatic carcinoma SW1990 cells

Chen Ruidong, Tang Wen, Hu Duanmin

Department of Gastroenterology, Second Affiliated Hospital of Soochow University, Suzhou 215004, China

Online:2015-07-08 Published:2015-05-26
Contact: Hu Duanmin E-mail:huduanmin@163.com

摘要/Abstract

摘要： 目的构建人类母源性印记基因3（MEG3）的真核表达载体，转染得到过表达的人胰腺癌SW1990细胞株，分析MEG3过表达对SW1990细胞增殖的影响。方法根据GenBank中MEG3的基因序列，通过化学合成的方法进行全基因合成得到人MEG3基因全长，将其构建入pcDNA3.0真核表达载体中，得到pcDNA3.0MEG3表达载体，转染胰腺癌细胞SW1990。采用RTPCR及荧光定量PCR技术检测转染细胞中MEG3基因的表达量。采用MTT试剂盒对转染后SW1990细胞增殖能力的变化进行检测。实验中设转染pcDNA3.0的SW1990细胞为阴性对照组，普通SW1990细胞为空白对照组。结果成功构建了MEG3真核表达质粒pcDNA3.0MEG3，并成功转染SW1990细胞。与两对照组相比，转染后细胞的MEG3基因表达量显著提高，提高约895倍（F=73.592，P<0.01）。MTT检测结果显示，MEG3基因明显抑制SW1990细胞的增殖，实验组72 h时吸光度值为0.81±0.06，与阴性对照组（1.17±0.07）和空白对照组(1.08±0.03）相比，3组差异有统计学意义（F=33.489，P<0.01）。结论本研究成功构建了MEG3真核表达质粒pcDNA3.0MEG3，并证实MEG3基因及其产物对胰腺癌SW1990细胞增殖有明显的抑制作用。

关键词: 胰腺肿瘤, RNA, 基因表达调控, 母源性印记基因3

Abstract: ObjectiveTo construct a maternally expressed gene 3 (MEG3) expression plasmid vector, and to obtain MEG3 overexpressed human pancreatic carcinoma SW1990 cells by transfection, and to analyze the effect of MEG3 overexpression on the proliferation of human pancreatic carcinoma SW1990 cells. MethodsA complete gene sequence based on the sequence of MEG3 in the GenBank was designed and inserted into the eukaryotic expression vector pcDNA3.0 to construct recombinant plasmid pcDNA3.0MEG3. It was identified by sequencing and transfected into human pancreatic carcinoma SW1990 cells. The expression of MEG3 in SW1990 cells was confirmed by RTPCR. The effect of MEG3 on proliferation was evaluated by MTT assay. In this study, the SW1990 cells transfected by plasmid pcDNA3.0 were named negative control group, and the usual SW1990 cells were named blank control group. ResultsA MEG3 expression plasmid vectorpcDNA3.0MEG3 was constructed successfully. And pcDNA3.0MEG3 vector was transfected into SW1990 cells successfully. The expression of MEG3 at mRNA in MEG3SW1990 cells increased significantly, about 895 times (F=73.592， P<0.01). The results of MTT assay indicated that overexpressed MEG3 could obviously inhibit SW1990 cells proliferation in vitro. After SW1990 cells transfected with pcDNA3.0MEG3 for 72 hours, the absorbance value was 0.81±0.06, with a statistically significance (F=33.489, P<0.01) compared with negative control group (1.17±0.07) and blank control group (1.08±0.03). ConclusionA MEG3 expression plasmid vectorpcDNA3.0MEG3 is constructed successfully. It is confirmed that MEG3 and its product have obvious inhibitory effects for the proliferation of human pancreatic carcinoma SW1990 cells.

Key words: Pancreatic neoplasms, RNA, Gene expression regulation, Maternally expressed gene 3

陈瑞东,唐文,胡端敏. 人MEG3基因质粒载体的构建及其对人胰腺癌SW1990细胞增殖的影响[J]. 国际肿瘤学杂志, 2015, 42(7): 488-491.

Chen Ruidong, Tang Wen, Hu Duanmin. Construction and inhibitory effect of MEG3 expression plasmid vector on the proliferation of human pancreatic carcinoma SW1990 cells[J]. Journal of International Oncology, 2015, 42(7): 488-491.

参考文献

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人MEG3基因质粒载体的构建及其对人胰腺癌SW1990细胞增殖的影响

Construction and inhibitory effect of MEG3 expression plasmid vector on the proliferation of human pancreatic carcinoma SW1990 cells

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