Shinoda Shuhei, Kaino Seiji, Amano Shogo, Harima Hirofumi, Matsumoto Toshihiko, Fujisawa Koichi, Takami Taro, Yamamoto Naoki, Yamasaki Takahiro, Sakaida Isao
Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan.
Department of Oncology and Laboratory Medicine, Yamaguchi University, Graduate School of Medicine, Yamaguchi, Japan.
Oncotarget. 2018 Jun 19;9(47):28434-28444. doi: 10.18632/oncotarget.25421.
Iron is an essential element for cell proliferation and growth processes. We have reported that deferasirox (DFX), an oral iron chelator, showed antiproliferative activity against pancreatic cancer cells. This study aimed to elucidate the effects of combination of gemcitabine (GEM), standard chemotherapy for pancreatic cancer, and DFX and .
GEM+DFX showed antiproliferative activity and induced apoptosis in pancreatic cancer cells . GEM+DFX suppressed xenograft tumor growth and induced apoptosis without any serious side effects compared with control, GEM, and DFX (average tumor volume: control 697 mm vs GEM 372 mm, < 0.05; GEM 372 mm vs GEM+DFX 234 mm, < 0.05). RRM1 and RRM2 protein levels were substantially reduced by DFX in BxPC-3 .
GEM+DFX has significant anticancer effects on pancreatic cancer cell through RR activity suppression.
BxPC-3, a human pancreatic cancer cell line, was used in all experiments. Cellular proliferation rate was measured using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt assay. Apoptosis was evaluated by flow cytometry and by measuring caspase 3/7 activity with luminescence assay. In the tumor xenografts in nude mice models, when five weeks after engraftment, drug administration began (day 0). After treatment for 21 days, the mice were sacrificed and the tumors were excised. Apoptotic cells in xenografts were evaluated by terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling assay. Protein levels of ribonucleotide reductase (RR) subunit 1 (RRM1) and RR subunit 2 (RRM2) in BxPC-3 cells were assessed by western blot .
铁是细胞增殖和生长过程中的必需元素。我们曾报道，口服铁螯合剂地拉罗司（DFX）对胰腺癌细胞具有抗增殖活性。本研究旨在阐明胰腺癌的标准化疗药物吉西他滨（GEM）与DFX联合使用的效果。
GEM+DFX对胰腺癌细胞具有抗增殖活性并诱导其凋亡。与对照组、GEM组和DFX组相比，GEM+DFX抑制异种移植瘤生长并诱导凋亡，且无任何严重副作用（平均肿瘤体积：对照组697 mm³，GEM组372 mm³，P<0.05；GEM组372 mm³，GEM+DFX组234 mm³，P<0.05）。在BxPC-3细胞中，DFX使RRM1和RRM2蛋白水平大幅降低。
GEM+DFX通过抑制RR活性对胰腺癌细胞具有显著抗癌作用。
所有实验均使用人胰腺癌细胞系BxPC-3。采用3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑内盐法测定细胞增殖率。通过流式细胞术和用发光法测量caspase 3/7活性评估凋亡情况。在裸鼠模型的肿瘤异种移植实验中，接种后五周开始给药（第0天）。治疗21天后，处死小鼠并切除肿瘤。通过末端脱氧核苷酸转移酶脱氧尿苷三磷酸缺口末端标记法评估异种移植瘤中的凋亡细胞。通过蛋白质印迹法评估BxPC-3细胞中核糖核苷酸还原酶（RR）亚基1（RRM1）和RR亚基2（RRM2）的蛋白水平。
