反义缺氧诱导因子1α对胰腺癌进展、转移及化疗敏感性的影响

Effect of antisense hypoxia-inducible factor 1alpha on progression, metastasis, and chemosensitivity of pancreatic cancer.

作者信息

Chang Qing, Qin Renyi, Huang Tao, Gao Jun, Feng Yanping

机构信息

Department of Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.

出版信息

Pancreas. 2006 Apr;32(3):297-305. doi: 10.1097/00006676-200604000-00010.

DOI:10.1097/00006676-200604000-00010

PMID:16628086

Abstract

OBJECTIVES

The aim of the study was to observe the effect of antisense hypoxia-inducible factor 1alpha (HIF-1alpha) on progression, metastasis, and chemosensitivity of pancreatic cancer.

METHODS

BxPc-3 cells transfected with antisense HIF-1alpha plasmid were exposed to 0.5% O2 for 4 hours. Expressions of HIF-1alpha, survivin, and beta1 integrin were detected by reverse transcriptase -polymerase chain reaction and Western blotting. Growth inhibition rates and apoptosis rates of BxPc-3 cells under different dosages of chemotherapy agents (5-fluorouracil, doxorubicin, and gemcitabine) were measured by MTT colorimetric assay and flow cytometry. The migration of BxPc-3 cells was assayed using transwell cell culture chambers. Subcutaneous transplantation of BxPc-3 cells in nude mice for 8 weeks was to assess progression and metastasis of pancreatic cancer.

RESULTS

Expression of HIF-1alpha was obviously down-regulated, and at the same time, survivin and beta1-integrin expressions were markedly down-regulated in the experimental group (P < 0.05). Higher dosages (100, 200, and 400 mg/L of 5-fluorouracil; 0.05, 0.075, and 0.1 mg/L of doxorubicin; and 10(-9), 10(-8), and 10(-7) mol/L of gemcitabine) caused a greater increase of inhibition in the experimental group than in control (P < 0.05). The number of migrated BxPc-3 cells in the experimental group was far less than in control (P < 0.05). In vivo, the tumor size and weight in the experimental group were significantly lower than those in control (P < 0.05).

CONCLUSION

Our data demonstrate that antisense HIF-1alpha inhibits expressions of survivin and beta1 integrin, enhancing apoptosis in human pancreatic cancer cells and restraining the progression and metastasis of pancreatic cancer. Therefore, HIF-1alpha may play a very important role in progression, metastasis, and chemosensitivity of human pancreatic cancer. Blocking HIF-1alpha in pancreatic cancer cells may offer an avenue for gene therapy.

摘要

目的

本研究旨在观察反义缺氧诱导因子1α（HIF-1α）对胰腺癌进展、转移及化疗敏感性的影响。

方法

将转染反义HIF-1α质粒的BxPc-3细胞置于0.5%氧气环境中4小时。采用逆转录-聚合酶链反应和蛋白质免疫印迹法检测HIF-1α、生存素及β1整合素的表达。通过MTT比色法和流式细胞术检测不同剂量化疗药物（5-氟尿嘧啶、阿霉素和吉西他滨）作用下BxPc-3细胞的生长抑制率和凋亡率。使用Transwell细胞培养小室检测BxPc-3细胞的迁移情况。将BxPc-3细胞皮下接种于裸鼠8周，以评估胰腺癌的进展和转移情况。

结果

实验组中HIF-1α的表达明显下调，同时生存素和β1整合素的表达也显著下调（P<0.05）。较高剂量（5-氟尿嘧啶100、200和400mg/L；阿霉素0.05、0.075和0.1mg/L；吉西他滨10^(-9)、10^(-8)和10^(-7)mol/L）时，实验组的抑制作用增强，与对照组相比差异有统计学意义（P<0.05）。实验组中迁移的BxPc-3细胞数量远少于对照组（P<0.05）。在体内，实验组的肿瘤大小和重量均显著低于对照组（P<0.05）。