摘要
目的观察转染人Endostatin(hEndostatin)基因对胰腺癌细胞株SW1990增殖、凋亡及血管内皮生长因子(VEGF)表达的影响。方法携带hEndostatin基因的复制缺陷型重组腺病毒载体体外转染SW1990细胞,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,实时荧光定量PCR和ELISA分析检测hEndostatin和VEGF的表达。建立裸鼠胰腺癌模型,随机分为3组(n=8),瘤体内分别注射磷酸盐缓冲液(PBS组)、报告基因LacZ重组腺病毒(Ad-LacZ组)、hEndostatin重组腺病毒(Ad-hEnd组)100μl,隔日1次,共4次。4周后免疫组织化学染色检测肿瘤组织中hEn- dostatin、VEGF、增殖性细胞核抗原(PCNA)的表达,TUNEL法检测肿瘤细胞凋亡。结果体外转染hEndostatin基因不影响SW1990细胞的增殖和凋亡(P>0.05),但下调VEGF的表达(P<0.01),下调作用第5天最大,在mRNA和蛋白水平的抑制率分别为84.67%和81.41%。体内转染4周后,Ad- hEnd组VEGF、PCNA阳性表达率分别为(36.3±7.1)%、(38.2±3.9)%,显著低于Ad-LacZ组的(81.2±6.6)%、(93.2±4.9)%和PBS组的(78.4±6.2)%、(90.1±5.7)%(P<0.01);肿瘤细胞凋亡率为(31.2±5.4)%,显著高于Ad-LacZ组的(9.4±4.9)%和PBS组的(8.5±3.7)%(P<0.01)。结论hEndostatin基因转染抑制SW1990细胞的体内增殖并促进凋亡;下调SW1990细胞内源性VEGF的表达可能是抗肿瘤的作用机制之一。
Objective To oberserve the effect of human endostatin gene transfection on cell proliferation ,apoptosis and the expression of vascular endothelial factor （VEGF） in SW1990 cell line. Methods SW1990 cells were infected by replication defective adenovirus vector containing human endostatin gene cDNA in vitro,then cell proliferation, apoptosis, expression of endostatin and VEGF were examined. Animal models of pancreatic carcinoma bearing nude mice were established by subcutaneous injection of SW1990 cells and randomized into 3 groups：Ad-hEnd group, Ad-LacZ group and PBS group. All animals were treated once 2 days with 4 times. The expression of endostatin, VEGF and proliferating cell nuclear antigen （PCNA） was detected by immtmohistochemistral staining and cell apoptosis by TUMEL in situ respectively after 4 weeks. Results Human endostatin gene had no effects on the proliferation and apoptosis of SW1990 cell line in vitro ,but down-regulated the expression of VEGF significantly with maximal inhibitory rate on 5th day at mRNA level （84.67%） and protein level （81.41%）. Positive rate of VEGF and PCNA in Ad-hEnd group [ （ 36.3 ± 7.1 ） % and （ 38.2 ± 3.9%, respectively] was significantly lower than that in Ad-LacZ group [（81.2±6.6）% and （93.2±4.9）% ,respectively） and PBS group [（78.4± 6.2） % and （90.1± 5.7 ） %, respectively ] （ P 〈 0.01 ）. Tumor cell apoptotic rate in Ad-hEnd group was （31.2 ± 5.4） %, significantly higher than that in Ad-LacZ group （9.4 ± 4.9） % and PBS group （ 8.5 ± 3.7 ）% （P 〈 0.01 ）. There was no significant difference between Ad-LacZ group and PBS group. Conclusion Human endostatin gene transfection inhibits SW1990 cell proliferation and promotes apeptosis in vivo. The down-regulation of the VEGF expression perhaps is one of its anti-tumor mechanisms.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第7期826-828,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(30200275)
关键词
内皮抑素
血管内皮生长因子
胰腺肿瘤
基因治疗
Endostatin
Vascular endothelial growth fator
Pancreatic neoplasms
Gene therapy
