[干扰素α对人肝癌细胞系及异种移植瘤中胸苷磷酸化酶的上调作用与抗血管生成]
[Up-regulation of thymidine phosphorylase and anti-angiogenesis by interferon alpha in human hepatocellular carcinoma cell line and xenograft].
作者信息
Xiao Yong-sheng, Zhou Jian, Tang Zhao-you, Fan Jia, Wu Zhi-quan, Liu Yin-kun, Ye Sheng-long, Shen Zao-zhuo, Xue Qiong, Zhao Yan
机构信息
Liver Cancer Institute and Zhongshan Hospital, Fudan University, Shanghai 200032, China.
出版信息
Zhonghua Yi Xue Za Zhi. 2005 Nov 30;85(45):3205-9.
OBJECTIVE
To further study the impact of interferon-alpha (IFN-alpha) on thymidine phosphorylase (TP) expression and angiogenesis.
METHODS
Human hepatocellular carcinoma cells of the line SMMC-7721 were cultured and added with IFN-alpha of different doses: 0 (as control group), 1000, 5000 and 10,000 U/ml. Twenty-four hours later RT-PCR was used to detect the TP mRNA expression. Boyden chamber method was used to examine the endothelial cells migration. Suspension of SMMC-7721 cells was inoculated subcutaneously into 30 male BALB/c-nu/nu mice, the mice were randomly divided into 5 equal groups to be subcutaneously injected with IFN-alpha of different doses: 0 (as control group), 1.0 x 10(6), 3.0 x 10(6), 9.0 x 10(6), and 1.5 x 10(7) U.kg(-1).d(-1) for 3 weeks. The eating behavior, activity, body weight, and tumor size were observed. The rats were killed 2 days after the drug injection was stopped. The subcutaneous tumors were taken out to undergo histological examination and TP protein expression by ELISA. The microvessel density (MVD) was detected using anti-CD34 monoclonal antibody.
RESULTS
The TP mRNA expression of the SMMC-7721 cells induced by IFN-alpha of the doses of 5000 U/ml and 10,000 U/ml significantly increased in comparison with the un-treated SMMC-7721 cells (0 U/ml, P < 0.05). The endothelial cell migration significantly increased in the IFN-alpha 1000 U/ml group compared with the control group (P < 0.001), and then decreased along with the increase of IFN-alpha dose; there were no significant differences in the epithelial migration among the groups of 0, 5000, and 10,000 U/ml IFN-alpha doses (all P > 0.05). The TP protein expression levels of the tumor in the rats treated with IFN-alpha of the doses of 9.0 x 10(6), and 1.5 x 10(7) U.kg(-1).d(-1) were 48 ng/mg +/- 24 ng/mg and 60 ng/mg +/- 6 ng/mg respectively, 1.9 and 2.4 times that of the control group (both P < 0.01). The MVD of the tumors was 6.0 +/- 1.8 in the 9.0 x 10(6) U.kg(-1).d(-1) IFN-alpha group, significantly higher than that of the control group (P < 0.01); and was 4.0 +/- 1.5 in the 1.5 x 10(7) U.kg(-1).d(-1) group, significantly lower than that of the 9.0 x 10(6) U.kg(-1).d(-1) group (P < 0.05) and not significantly different from that of the control group. The tumor inhibiting rate of the 1.5 x 10(7) U.kg(-1).d(-1) IFN-alpha group was 68%, significantly higher than that of the untreated group (P < 0.05).
CONCLUSION
IFN-alpha of certain doses up-regulate the TP expression, and inhibit the angiogenesis induced by TP as well.
目的
进一步研究α-干扰素（IFN-α）对胸苷磷酸化酶（TP）表达及血管生成的影响。
方法
培养人肝癌SMMC-7721细胞株，分别加入不同剂量的IFN-α：0（作为对照组）、1000、5000和10000 U/ml。24小时后采用逆转录-聚合酶链反应（RT-PCR）检测TP mRNA表达。采用Boyden小室法检测内皮细胞迁移。将SMMC-7721细胞悬液皮下接种于30只雄性BALB/c-nu/nu小鼠，将小鼠随机分为5组，每组6只，分别皮下注射不同剂量的IFN-α：0（作为对照组）、1.0×10⁶、3.0×10⁶、9.0×10⁶和1.5×10⁷ U·kg⁻¹·d⁻¹，共3周。观察进食行为、活动、体重及肿瘤大小。停药2天后处死大鼠。取出皮下肿瘤进行组织学检查，并用酶联免疫吸附测定（ELISA）法检测TP蛋白表达。用抗CD34单克隆抗体检测微血管密度（MVD）。
结果
与未处理的SMMC-7721细胞（0 U/ml）相比，5000 U/ml和10000 U/ml剂量的IFN-α诱导的SMMC-7721细胞TP mRNA表达显著增加（P<0.05）。与对照组相比，1000 U/ml IFN-α组内皮细胞迁移显著增加（P<0.001），随后随着IFN-α剂量的增加而降低；0、5000和10000 U/ml IFN-α剂量组之间上皮迁移无显著差异（均P>0.05）。9.0×10⁶和1.5×10⁷ U·kg⁻¹·d⁻¹剂量的IFN-α处理的大鼠肿瘤中TP蛋白表达水平分别为48 ng/mg±24 ng/mg和60 ng/mg±6 ng/mg，分别是对照组的1.9倍和2.4倍（均P<0.01）。9.0×10⁶ U·kg⁻¹·d⁻¹ IFN-α组肿瘤的MVD为6.0±1.8，显著高于对照组（P<0.01）；1.5×10⁷ U·kg⁻¹·d⁻¹组为4.0±1.5，显著低于9.0×10⁶ U·kg⁻¹·d⁻¹组（P<0.05），与对照组无显著差异。1.5×10⁷ U·kg⁻¹·d⁻¹ IFN-α组的肿瘤抑制率为68%，显著高于未处理组（P<0.05）。
结论
一定剂量的IFN-α上调TP表达，并抑制由TP诱导产生的血管生成。
Zotero 插件