重组分泌型肿瘤坏死因子相关凋亡诱导配体腺病毒对PC-3细胞荷瘤鼠瘤体的生长抑制

Suppression effect of secreting-trail adenovirus on the prostate cancer PC-3 cells in the tumor bearing mice

目的探讨表达分泌型TRAIL基因的腺病毒载体对人前列腺癌PC-3细胞荷瘤鼠的生长抑制作用。方法 DAPI染色检测各组PC-3细胞凋亡的情况。Transwell实验检测Ad-s TRAIL对PC-3细胞体外侵袭能力的影响。成功建立PC-3荷瘤鼠模型,经Ad-s TRAIL和Ad-Lac Z分别处理后检测瘤体的生长抑制率。免疫组化实验检测裸鼠移植瘤内TRAIL表达的变化。TUNEL检测瘤体内肿瘤细胞的凋亡。结果 DAPI染色显示Ad-s TRAIL组肿瘤细胞凋亡水平明显高于Control组和Ad-Lac Z组。Transwell实验显示Ad-s TRAIL组穿入下室面的细胞数(24.8±3.70)明显少于对照组(47.6±4.28,q=10.68,P<0.01)及Ad-Lac Z组(49.6±4.39,q=11.61,P<0.01)。成瘤实验表明:Ad-s TRAIL对荷瘤鼠肿瘤的抑制率3 d为8.15%,6 d为11.55%,9 d为17.23%,12 d为20.05%,15 d为27.18%,18 d为34.27%,21 d为33.08%。免疫组化实验显示:Ad-s TRAIL组荷瘤鼠肿瘤组织中TRAIL表达水平明显高于Control组和Ad-Lac Z组。TUNEL实验显示:Ad-s TRAIL组荷瘤鼠肿瘤细胞凋亡水平明显高于Control组和Ad-Lac Z组。结论通过重组分泌型TRAIL腺病毒载体Ad-s TRAIL处理PC-3细胞荷瘤鼠,可以抑制裸鼠体内肿瘤细胞的增殖,促进前列腺癌细胞的凋亡。

Objective To investigate the suppression effect of secreting-trail adenovirus on the prostatecancer PC-3 cells in the tumor bearing mice. Methods Apoptosis of PC-3 cells was measured by DAPI staining in vitro. Invasion of PC-3 cells was measured by transwell chamber assay. The tumor bearing mice were prepared andused to test the inhibition rates in different groups. The protein expression of TRAIL in different groups tumorbearing mice was determined by immunohistochemistry. The variation of cell apoptosis in different groups of tumorbearing mice was detected by TUNEL assay. Results DAPI staining showed that the apoptosis level in Ad-sTRAILgroup was significantly higher than that in the control group and that in the Ad-Lac Z group,respectively. Thenumber of PC-3 cells invading the inferior chamber in the Ad-sTRAIL group（24.8 ± 3.70）was significantlydecreased compared with that in the control group（47.6 ± 4.28,q = 10.68,P〈0.01）and that in the Ad-Lac Z group（49.6 ± 4.39,q = 11.61,P〈0.01）. Growth inhibition assays in vivo showed the inhibition rate of Ad-sTRAIL was3 d 8.15%,6 d 11.55%,9 d 17.23%,12 d 20.05%,15 d 27.18%,18 d 34.27%,and 21 d 33.08%,respective-ly. Immunohistochemistry results showed that the expression level of TRAIL in tumors of Ad-sTRAIL group tumor-bearing mice was significantly higher than that in the control group and that in the Ad-Lac Z group. TUNEL assayshowed that the apoptosis level of tumor bearing mice tumor cells in the Ad-sTRAIL group was significantly higherthan that in the control group and that in the Ad-Lac Z group. Conclusion The secreting-trail adenoviral vectorcould inhibit the growth of tumor bearing mice tumor cells,furthermore it could induce the apoptosis of PC-3 cellsin the tumor bearing mice.