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通过构建融合RGD肽的NF-κB抑制肽,获取新型抗瘤融合肽,研究其抗肿瘤效果。构建人造抗瘤活性肽p EGFP-C1-ARP表达载体,转染COS7细胞,获取人造抗瘤活性肽,利用Western blotting法检测PC-3M细胞A20、Bcl2、CCND1;通过观察ARP对荷人前列腺癌PC-3M裸鼠肿瘤抑制实验,研究人造抗瘤活性肽的抗肿瘤效果。限制性内切酶酶切鉴定与DNA测序证实合成人造抗瘤活性肽真核载体寡核苷酸链插入正确。Western blotting检测结果表明转染后PC-3M细胞A20蛋白表达增加。并抑制凋亡抑制基因Bcl2的表达,动物实验证实人造抗瘤活性肽可显著抑制肿瘤生长。本研究成功构建人造抗瘤活性肽,其对肿瘤细胞具有明显的消除效应。
The anti-tumor effect of a novel anti-tumor fusion peptide was obtained by constructing an NF-κB-inhibiting peptide fused with RGD peptide. The antitumor active peptide p EGFP-C1-ARP expression vector was constructed and transfected into COS7 cells to obtain artificial antitumor active peptides. Western blotting was used to detect the expression of A20, Bcl2 and CCND1 in PC-3M cells. PC-3M tumor inhibition in nude mice to study the antitumor effect of artificial antitumor peptides. Restriction endonuclease digestion and DNA sequencing confirmed that the synthetic antitumor active peptide eukaryotic vector oligonucleotide insert correctly. Western blotting results showed that A20 protein expression in PC-3M cells increased after transfection. And inhibit the expression of apoptosis-inhibiting gene Bcl2. Animal experiments confirmed that the artificial anti-tumor active peptide can significantly inhibit tumor growth. This study successfully constructed artificial anti-tumor active peptide, which has obvious elimination effect on tumor cells.