Construction of DPC4 Gene Recombinant Expression Vector and Its Inhibitory Effects on Human Pancreatic Adenocarcinoma Cell Line Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

YUAN Shengguang ¹ , TIAN Ling ² , WEI Yuquan ² , et al. ¹

Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041;

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Keywords：

DPC4 gene Pancreas adenocarcinoma Cell cycle

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ObjectiveTo construct DPC4 gene recombinant expression vector and to study the inhibitory effect of DPC4 on the growth of human pancreatic adenocarcinoma cell line (PC3) cells.MethodsDPC4 cDNA was amplified from K562 cell line using RTPCR, and was cloned into the pcDNA3.1 vector to construct a recombinant expression vector plasmid pcDNA3.1DPC4. The recombinant expression plasmid was transferred into PC3 cells by liposome method. After G418 selection, cell cycle and apoptosis were assessed by flow cytometry, then the cell growth rate was estimated by cell count. The cells being not transferred plasmid and transferred pcDNA3.1 plasmid were used as controls.ResultsThe DPC4 gene recombinant expression vector was constructed. Wildtype DPC4 gene attributed to the increase of G1 phase cells and the decrease of S phase cells in PC3 cells,and could inhibit the growth of PC3 cells, the cell growth rates was reduced to 34.3%-41.1% of that of the controls, but cell apoptosis was not observed on all groups. ConclusionWildtype DPC4 gene could inhibit the proliferation of human pancreatic adenocarcinoma cells and could become one of the target genes of pancreas adenocarcinoma gene therapy

Citation： YUAN Shengguang,TIAN Ling,WEI Yuquan,et al.. Construction of DPC4 Gene Recombinant Expression Vector and Its Inhibitory Effects on Human Pancreatic Adenocarcinoma Cell Line Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2002, 9(6): 374-376. doi: Copy

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1. Duff EK, Clarke AR. Smad4(DPC4)a potent tumour suppressor？ [Ｊ]. Br J Cancer, 1998; 78(12)∶1615.
2. 卢圣栋主编. 现代分子生物学实验技术 [M]. 第2版. 北京：中国协和医科大学出版社， 1999∶233～658.
3. F奥斯伯， R布伦特， RE 金斯，等著. 颜子颖，王海林译. 精编分子生物学实验指南 [M]. 第1版. 北京：科学出版社，1998∶ 1～125.
4. Grau AM, Zhang L, Wang W, et al. Induction of p21wafl expression and growth inhibition by transforming growth factor β involve the tumor suppressor gene DPC4 by human pancreatic adenocarcinoma cells [Ｊ]. Cancer Res, 1997; 57(18)∶3929.
5. Dai JL, Bansal RK, Kern SE. G1 cell cycle arrest and apoptosis induction by nuclear Smad4/DPC4: phenotypes reversed by a tumorigenic mutation [Ｊ]. Proc Natl Acad Sci USA, 1999; 96(4)∶1427.
6. 王忠，严律南，李立，等. 细胞周期相关调控蛋白在胰腺癌中的表达及其意义 [Ｊ]. 中国普外基础与临床杂志， 2000； 7（5）∶279.
7. Hunt KK, Fleming JB, Abramian A, et al. Overexpression of the tumor suppressor gene Smad4/DPC4 induces p21wafl expression and growth inhibition in human carcinoma cells [Ｊ]. Cancer Res, 1998; 58(24)∶5656.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Construction of DPC4 Gene Recombinant Expression Vector and Its Inhibitory Effects on Human Pancreatic Adenocarcinoma Cell Line Cells

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