Department of Hepatobiliary Surgery, Ningxia Medical University, Yinchuan 750004, Ningxia, China.
Int J Mol Sci. 2013 Dec 13;14(12):24293-304. doi: 10.3390/ijms141224293.
The influence of Glucagon-like peptide-1 (GLP-1) and Exendin-4 on development of intrahepatic cholangiocarcinoma (ICC) is evaluated in the study. In vitro tests, including acute toxicity test, cell colony formation assays, cells proliferation and apoptosis, transwell assay, were performed. An ICC in situ tumor animal model was established. Then, animals were randomly divided into four groups (n = 6): control, Exendin-4 treatment, oxaliplatin treatment and Exendin-4-oxaliplatin treatment. Animals in the Exendin-4 treatment and Exendin-4-oxaliplatin treatment groups received a subcutaneous injection of Exendin-4 (100 μg/kg/day) for 1 week, and then received oxaliplatin (10 mg/kg/week) by tail vein injection. Animals in the control group received PBS. Immunohistochemistry tests were used for PCNA, Ki67, Caspase 3 expression in tumor tissue. Results show that that, after incubation of human cholangiocarcinoma cell lines, HuCCTI and GLP-1, or HuCCTI and Exendin-4, colony formation number was sharply decreased. However, GLP-1, HuCCTI or Exendin-4 did not affect the colony of normal cells. Combination treatment with oxaliplatin and Exendin-4 can significantly inhibit tumor cells' proliferation and promote apoptosis. The combined effect is stronger than that of oxaliplatin or Exendin-4. Combination treatment with oxaliplatin and Exendin4 can significantly decrease Ki67 and PCNA proteins' expression in subcutaneous tumors of nude mice. The inhibitory effect of Combination treatment with oxaliplatin and Exendin4 is clearly stronger than that of oxaliplatin. In addition, Combination treatment with oxaliplatin and Exendin4 can significantly increase Caspase3 protein positive expression. In short, these results show that combination treatment with oxaliplatin and Exendin4 can inhibit tumor cells' proliferation, and promote apoptosis.
本研究评估了胰高血糖素样肽-1（GLP-1）和 Exendin-4 对肝内胆管癌（ICC）发展的影响。进行了体外试验，包括急性毒性试验、细胞集落形成试验、细胞增殖和凋亡试验、Transwell 试验。建立了 ICC 原位肿瘤动物模型。然后，动物随机分为四组（n = 6）：对照组、Exendin-4 治疗组、奥沙利铂治疗组和 Exendin-4-奥沙利铂治疗组。Exendin-4 治疗组和 Exendin-4-奥沙利铂治疗组的动物接受皮下注射 Exendin-4（100μg/kg/天）1 周，然后通过尾静脉注射奥沙利铂（10mg/kg/周）。对照组动物给予 PBS。免疫组织化学试验用于检测肿瘤组织中 PCNA、Ki67、Caspase 3 的表达。结果表明，人胆管癌细胞系 HuCCTI 与 GLP-1 或 HuCCTI 与 Exendin-4 孵育后，集落形成数明显减少。然而，GLP-1、HuCCTI 或 Exendin-4 对正常细胞的集落没有影响。奥沙利铂和 Exendin-4 联合治疗可显著抑制肿瘤细胞增殖并促进凋亡。联合作用强于奥沙利铂或 Exendin-4。奥沙利铂和 Exendin4 联合治疗可明显降低裸鼠皮下肿瘤中 Ki67 和 PCNA 蛋白的表达。奥沙利铂联合治疗的抑制作用明显强于奥沙利铂。此外，奥沙利铂和 Exendin4 联合治疗可显著增加 Caspase3 蛋白的阳性表达。总之，这些结果表明，奥沙利铂和 Exendin4 的联合治疗可以抑制肿瘤细胞的增殖，促进凋亡。
