血管内皮生长因子反义寡核苷酸对体外及裸鼠体内前列腺癌细胞PC3生物学特性影响的研究

 目的　探讨血管内皮生长因子（VEGF）反义寡核苷酸对前列腺癌细胞PC3在体外和裸鼠体内生长特性的影响，以及局部注射反义寡核苷酸治疗裸鼠皮下移植肿瘤的疗效。方法　采用Oligofectamine携带VEGF反义寡核苷酸转染前列腺癌细胞PC3，实验分为反义寡核苷酸组、正义寡核苷酸组和对照组。软琼脂糖凝胶实验和细胞侵袭实验检测转染反义寡核苷酸的肿瘤细胞成瘤性和侵袭性。裸鼠成瘤实验观察VEGF反义寡核苷酸对体内PC3细胞增殖的影响。建立裸鼠前列腺癌种植瘤模型，局部注射VEGF反义核酸治疗，测定体内抑瘤率。结果　对照组、正义寡核苷酸组和反义寡核苷酸组的PC3细胞每组阳性克隆数分别为53.67±5.86、52.33±6.43和26±4.58（F＝13.73，P＜0.01），反义组体外形成克隆数显著减少；三组细胞侵袭至下室的细胞数分别为45.60±5.53、42.35±6.21和18.37±3.52（F＝14.18，P＜0.01）；转染VEGF反义核酸的细胞移植瘤生长较慢，接种28 d后三组的肿瘤体积分别为（1330.32±81.38）、（1267.64±120.26）和（641.83±58.34）mm3（F＝17.26，P＜0.01）；局部注射治疗4周后，三组肿瘤质量分别为（1.25±0.08）g、（1.17±0.06）g和（0.41±0.05）g，与对照组比较，正义组和反义组肿瘤抑制率分别为6.4 %和67.2 %，差异有统计学意义（χ2＝17.72，P＜0.005）。结论　VEGF反义寡核苷酸可以抑制前列腺癌细胞PC3 VEGF的表达，进而促进细胞凋亡，降低其成瘤性，抑制侵袭能力。转染VEGF反义寡核苷酸的前列腺癌细胞PC3移植瘤在裸鼠体内生长受抑制，局部注射反义核酸可显著抑制移植瘤的生长。

Effect of VEGF ASODN in vitro and in vivo of nude mice on the biological characteristics of human prostate cancer PC3 cells

Objective To investigate the effect of VEGF ASODN in vitro and in vivo on the biological characteristics of human prostate cancer PC3 cells and its effect in xenotransplanted tumors in nude mice by local ASODN injection.Methods VEGF ASODN was delivered into PC3 cells by Oligofectamine.There were three experimental groups: VEGF ASODN,VEGF ODN and control.Soft agar assay and matrigel invasion assay were used to measure cellular transformation and invasion ability,respectively.Tumor formation assay in nude mice was used to evaluate the effect of VEGF ASODN on proliferation of PC3 cells in vivo.The xenotransplanted prostate tumor model in nude mice was established and the effect of local ASODN injection on the inhibition of tumor growth in vivo was examed.Results The soft agar colony numbers for control,ODN,and ASODN treated cells were 53.67±5.86,52.33±6.43 and 26.00±4.58,respectively (F =13.73,P＜0.01).The numbers of invaded cells for three group were 45.60±5.53,42.35±6.21 and 18.37±3.52,respectively (F =14.18,P ＜0.01).Tumor cells transfected with VEGF ASODN proliferated more slowly than other groups.28 days later after tumor cells were injected into nude mice,the tumor sizes of three groups were (1330.32±81.38) mm3,(1267.64±120.26) mm3 and (641.83±58.34) mm3 (F =17.26,P ＜0.01).After treating the transplanted tumor with VEGF ASODN or control oligos for four weeks,the tumor weight of three groups was (1.25±0.08) g,(1.17±0.06) g and (0.41±0.05) g,respectively.Comparing with control groups,the tumor inhibitory rates of ODN group and ASODN group were 6.4 ％ and 67.2 ％,respectively (x2=17.72,P＜0.005).Conclusion VEGF ASODN could inhibit VEGF expression in PC3 cells and lead to increasing cell apoptosis.After VEGF ASODN treatment,tumorigenesis in vitro is inhibited and cell invasion ability is decreased.The tumors originated from cells transfected with VEGF ASODN grow more slowly than control groups.Also local injection of VEGF ASODN could inhibit the growth of transplanted tumors in nude mice.