小鼠结肠癌CT26细胞PARP抑制后肝转移变化的影响

Effect of Poly (ADP-ribose) polymerase inhibition on liver metastasis of mouse colorectal carcinoma CT26 cell line in vivo

目的:探讨小鼠大肠癌CT26细胞聚腺苷二磷酸核糖聚合酶〔Poly(ADP-ribose)polymerase,PARP〕抑制后肝转移变化及其可能的机制.方法:采用小鼠结肠癌CT26细胞肝转移模型,5-氨基异喹啉酮(5-AIQ)为PARP抑制剂.实验组分为两组,分别腹腔注射5-AIQ3mg/(kg.d)和5-AIQ10mg/(kg.d),对照组给予相应体积双蒸水.14d后,观察各组小鼠脾脏原发肿瘤和肝脏转移肿瘤体积、数目的变化.WesternBlot检测PARP,NF-κB,基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)蛋白在各组脾脏原发瘤组织中的表达.结果:5-AIQ3mg/(kg.d)浓度组和5-AIQ10mg/(kg.d)浓度组脾脏肿瘤体积分别为(0.92±0.23)cm3和(0.88±0.43)cm3,与对照组(2.64±0.72)cm3相比较明显缩小,差异具有统计学意义(P<0.05);肝脏转移结节数目5-AIQ3mg/(kg.d)浓度组和5-AIQ10mg/(kg.d)浓度组分别为(4.20±2.95)个,(5.40±3.31)个,两组间差异无统计学意义(P>0.05),但与对照组(47.18±9.81)个相比较,差异均具有统计学意义(P<0.05).5-AIQ3mg/(kg.d)浓度组小鼠脾脏原发瘤组织PARP,NF-κB,MMP-2和MMP-9表达分别为(1.00±0.09),(0.55±0.03),(0.58±0.05),(0.48±0.06),与对照组(1.56±0.14),(0.84±0.07),(0.91±0.08)和(0.74±0.10)相比较明显降低,差异具统计学意义(P<0.05).结论:小鼠结肠癌CT26细胞PARP抑制可抑制肿瘤的生长及肝转移,可能与PARP通过NF-κB调节MMP-2,MMP-9蛋白表达有关.

AIM： To investigate the liver metastasis changes of CT26 cell line by Poly（ADP-ribose） polymerase （PARP） inhibition in vivo, and how it plays its role. METHODS： Liver metastasis model of colon carcinoma CT26 cell line was prepared in mice. 5-aminoisoquinolinone （5-AIQ）was used as the PARP inhibitor. After daily treatment with 5-AIQ 3mg/（kg · d） and 10 mg/（kg · d） was given ip. for 14 d, the mice were sacrificed by cervical dislocation. The mice in control group were given double distilled water instead. The number and size of the splenic implanted carcinoma amd liver metastatic carcinoma were observed and counted. Western Blot was used to detect the PARP, NF-κB, MMP-2 and MMP-9 expressions in spleen primary carcinoma. RESULTS ： The tumor volumes （ 0.92 ± 0.23 ） and （0.88 ±0.43）cm^3 in the treated groups were much smaller than （2.64 ± 0.72） cm^3 in the control group （ P 〈 0.05 ）. The number of liver metastatic nodules was（4.20 ± 2.95 ） , （5.40 ± 3.31 ） in the treated groups, less than（47.18 ±9.81 ） in the control group （P 〈 0. 05 ）, however, no statistical significance was found between the groups treated with different 5-AIQ doses （P 〉 0.05）. The expressions of PARP, NF-κB, MMP-2, MMP-9 in 3 mg/（kg · d） 5-AIQ treated group [ （ 1.00 ± 0.09）, （0.55 ± 0.03 ）, （0.58 ± 0.05 ） , （0.48 ± 0.06 ） ] were lower than those [ （1.56±0.14） ,（0.84 ±0.07） ,（0.91 ±0.08） ,（0.74 ±0.10） ] in control group, respectively （ P 〈 0.05 ） . CONCLUSION. PARP inhibition can inhibit the growth of primary splenic carcinoma and liver metastasis in vivo. It is probably correlated with the decreased expressions of NF-κB,MMP-2 and MMP-9 regulated by PARP.