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目的:构建携带Jagged 2(JAG2)基因siRNA的慢病毒表达载体,并观察其转染人胰腺癌细胞生物学特性的影响。方法:采用基因重组技术构建若干携JAG2基因siRNA片段的慢病毒表达载体,将其转染经酶解法从胰腺癌组织标本中原代分离的胰腺癌细胞;选择对JAG2基因抑制效率最高的siRNA片段,通过MTT法、流式细胞术、Transwell小室实验观察其转染对原代胰腺癌细胞生长、凋亡、细胞周期及侵袭转移能力的影响。结果:所构建的携JAG2基因siRNA片段的慢病毒表达载体均能不同程度降低原代胰腺癌细胞JAG2 mRNA与蛋白的表达。JAG2 siRNA转染后,胰腺癌细胞的增殖明显降低、凋亡与S期阻滞明显增强、侵袭转移能力明显减弱(均P<0.05)。结论:成功构建携JAG2基因siRNA慢病毒表达载体,其转染能有效削弱人胰腺癌细胞的恶性生物学特征。
OBJECTIVE: To construct a lentiviral vector carrying Jagged 2 (JAG2) gene siRNA and to observe its biological characteristics in transfecting human pancreatic cancer cells. METHODS: Several lentiviral expression vectors carrying JAG2 gene siRNA fragment were constructed by gene recombination technique and transfected into primary pancreatic cancer cells isolated from pancreatic cancer tissue samples by enzymatic method. The siRNA fragment with the highest JAG2 gene silencing efficiency was selected, The effects of transfection on the growth, apoptosis, cell cycle and invasion and metastasis of primary pancreatic cancer cells were observed by MTT assay, flow cytometry and Transwell chamber assay. Results: The constructed lentiviral vector carrying JAG2 gene siRNA fragment could reduce the expression of JAG2 mRNA and protein in primary pancreatic cancer cells to varying degrees. After transfection with JAG2 siRNA, the proliferation of pancreatic cancer cells was significantly decreased, and the arrest of apoptosis and S phase was significantly enhanced. The invasion and metastasis ability of JAG2 siRNA was significantly attenuated (all P <0.05). CONCLUSION: siRNA lentiviral vector carrying JAG2 gene has been successfully constructed. Its transfection can effectively attenuate the malignant biological characteristics of human pancreatic cancer cells.