5’-脱氧氟尿苷联合干扰素对转染TP基因结肠癌细胞株裸鼠实验模型的影响

发布时间：2018-02-07 12:07

本文关键词： 结直肠癌 LST 胸苷磷酸化酶 ’-脱氧氟尿苷干扰素-αa　出处：《中国癌症杂志》2017年11期 　论文类型：期刊论文

【摘要】：背景与目的:人结直肠癌细胞基本无胸苷磷酸化酶(thymidine phosphorylase,TP)表达。本实验主要探讨人结肠癌细胞株LS174T转染TP基因后,在裸鼠肿瘤组织中对抗癌药物5’-脱氧氟尿苷(5’-deoxy-5-fluorouridine,5’-DFUR)、氟尿嘧啶(5-fluorouracil,5-FU)和干扰素-α2a(interferon-α2a,INF-α2a)干预效果的影响。方法:以慢病毒为载体转染TP基因至结肠癌细胞株LS174T,得到稳定传代的高TP表达株。将96只BALB/c裸鼠分成2组,分别应用野生型及转染TP基因后的LS174T细胞株接种裸鼠背部皮下,制成结肠癌TP低、高表达模型。每组再随机分为6组,分别应用0.9%Na Cl溶液、INF-α2a、5-FU、5-FU+INF-α2a、5’-DFUR和5’-DFUR+INF-α2a干预5 d,2周后处死裸鼠,分离肿瘤称重后,分别应用鼠抗人CD34、血管内皮生长因子(vascular endothelial growth factor,VEGF)和抗PD-ECGF单克隆抗体进行免疫组织化学染色。判定是否有TP表达及对微血管进行计数。结果:在野生型LS174T裸鼠荷瘤模型中,使用抗肿瘤药物的3~6组肿瘤质量与对照组相比均明显降低,抑癌率分别为48%、27%、48%和57%,差异有统计学意义(P0.05)。其中5’-DFUR组(48%)与5’-DFUR+INF-α2a组(57%)、5-FU+INF-α2a组与5’-DFUR+INF-α2a组(27%与57%)相比,抑癌率差异有统计学意义(P0.05)。在LS174T-TP细胞株裸鼠模型中,5’-DFUR组和5’-DFUR+INF-α2a组抑癌率分别为27%和48%,而使用5-FU的两组未显示出良好的抗肿瘤效果(7%和3%)。免疫组织化学染色显示,野生型LS174T接种形成的肿瘤组织中TP表达阴性,微血管数目较少,而转染TP基因后,肿瘤组织中TP表达明显上升,微血管密度明显增多(P0.05)。结论:结肠癌细胞株LS174T转染TP基因后,在裸鼠实验肿瘤模型中,TP表达明显增加,使细胞内激活抗癌药物5’-DFUR的抗癌作用明显增强,联合应用INF-α2a能进一步增强这种抗癌作用。而转染TP基因或联合应用INF-α2a对常规抗癌药物5-FU的抗癌效果无明显影响。
[Abstract]:Background & AIM: there is no thymidine phosphorylase (TPTP) expression in human colorectal cancer cells. This study was designed to investigate the expression of TP gene in human colon cancer cell line LS174T. In nude mice tumor tissues, the effects of anti-cancer drugs 5- deoxy-5-fluorouridine5- deoxy-5-fluorouridine5, fluorouraciline 5-FU) and interferon-伪 2a interferon- 伪 2aF- 伪 2aF- 伪 2a) on the effects of anti-cancer drugs 5- deoxy-5-fluorouridine 5-fluorouraciline (5-FUU) and interferon-伪 2a interferon- 伪 2a- INF- 伪 2a) were obtained by transfection of TP gene into colon cancer cell line LS174T by lentivirus method: high TP was obtained by transfection of TP gene into colon cancer cell line LS174T. 96 BALB/c nude mice were divided into 2 groups. Nude mice were subcutaneously inoculated with wild type and LS174T cells transfected with TP gene to make a low and high expression model of TP in colon cancer. Each group was randomly divided into 6 groups. The nude mice were killed after 5 days of intervention with 0.9% NaCl solution (INF- 伪 2a) -FU 5-FU (5-FU) -5-FU (5-FU) -5-FU (5-FU) -5-FU (5-FU) -5-FU (5-FU). After separating the tumor and weighing it, Mouse anti-human CD34, vascular endothelial growth factor endothelial growth factor-VEGF and anti-#en2# monoclonal antibodies were used for immunohistochemical staining to determine the expression of TP and count the microvessels. Results: in the nude mice model of wild-type LS174T, the expression of TP was determined. Compared with the control group, the tumor quality of the three groups treated with antitumor drugs was significantly lower, and the tumor inhibition rates were 2748% and 57%, respectively. The difference was statistically significant (P 0.05). The difference was significant (P 0.05), and the tumor quality of the 5 + DFUR INF- 伪 2a group was significantly lower than that of the 55-FU INF- 伪 2a group (27% vs 5757%), and the tumor suppressor rate was 27% in the 5-DFUR INF- 伪 2a group and 57% in the 5- DFUR INF- 伪 2a group, respectively, compared with that in the 55-FUINF- 伪 2a group (P < 0.05). There was a significant difference in tumor suppressor rate between the two groups (P 0.05). In the LS174T-TP cell line nude mice model, the tumor inhibition rates of the groups were 27% and 48, respectively, while the two groups with 5-FU showed no good anti-tumor effect (7%) and 3Q (3%). Immunohistochemical staining showed that the tumor suppressor rate of the two groups was higher than that of the control group (P < 0.05), and that the tumor inhibition rates of the two groups were 27% and 48 respectively. TP expression was negative and the number of microvessels was small in tumor tissues inoculated with wild-type LS174T. After transfection of TP gene, TP expression and microvessel density in tumor tissues increased significantly (P 0.05). Conclusion: after transfection of TP gene into colon cancer cell line LS174T, TP gene expression is significantly increased. In nude mice tumor model, the expression of TP was increased, and the anti-cancer effect of the anti-cancer drug 5- DFUR was obviously enhanced. The combination of INF- 伪 2a and INF- 伪 2a could further enhance the anticancer effect, but transfection of TP gene or combined use of INF- 伪 2a had no significant effect on the anticancer effect of 5-FU.
【作者单位】：广州医科大学附属第二医院胃肠外科;
【基金】：国家自然科学基金(81071883)
【分类号】：R735.35

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