Kazim Sabiha, Malafa Mokenge P, Coppola Domenico, Husain Kazim, Zibadi Sherma, Kashyap Trinayan, Crochiere Marsha, Landesman Yosef, Rashal Tami, Sullivan Daniel M, Mahipal Amit
Department of Gastrointestinal Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida.
Department of Anatomic Pathology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida.
Mol Cancer Ther. 2015 Jul;14(7):1570-81. doi: 10.1158/1535-7163.MCT-15-0104. Epub 2015 May 1.
Pancreatic cancer is an aggressive and deadly malignancy responsible for the death of over 37,000 Americans each year. Gemcitabine-based therapy is the standard treatment for pancreatic cancer but has limited efficacy due to chemoresistance. In this study, we evaluated the in vitro and in vivo effects of gemcitabine combined with the selective nuclear export (CRM1) inhibitor KPT-330 on pancreatic cancer growth. Human pancreatic cancer MiaPaCa-2 and metastatic pancreatic cancer L3.6pl cell lines were treated with different concentrations of KPT-330 and gemcitabine alone or in combination, and anchorage-dependent/independent growth was recorded. In addition, L3.6pl cells with luciferase were injected orthotopically into the pancreas of athymic nude mice, which were treated with (i) vehicle (PBS 1 mL/kg i.p., 2/week and povidone/pluronic F68 1 mL/kg p.o., 3/week), (ii) KPT-330 (20 mg/kg p.o., 3/week), (iii) gemcitabine (100 mg/kg i.p., 2/week), or (iv) KPT-330 (10 mg/kg) + gemcitabine (50 mg/kg) for 4 weeks. KPT-330 and gemcitabine alone dose-dependently inhibited anchorage-dependent growth in vitro and tumor volume in vivo compared with vehicle treatment. However, the combination inhibited growth synergistically. In combination, KPT-330 and gemcitabine acted synergistically to enhance pancreatic cancer cell death greater than each single-agent therapy. Mechanistically, KPT-330 and gemcitabine promoted apoptosis, induced p27, depleted survivin, and inhibited accumulation of DNA repair proteins. Together, our data suggest that KPT-330 potentiates the antitumor activity of gemcitabine in human pancreatic cancer through inhibition of tumor growth, depletion of the antiapoptotic proteins, and induction of apoptosis.
胰腺癌是一种侵袭性强且致命的恶性肿瘤，每年导致超过37000名美国人死亡。基于吉西他滨的疗法是胰腺癌的标准治疗方法，但由于化疗耐药性，其疗效有限。在本研究中，我们评估了吉西他滨与选择性核输出（CRM1）抑制剂KPT-330联合使用对胰腺癌生长的体外和体内作用。用人胰腺癌MiaPaCa-2细胞系和转移性胰腺癌L3.6pl细胞系分别单独或联合使用不同浓度的KPT-330和吉西他滨进行处理，并记录贴壁依赖性/非依赖性生长情况。此外，将携带荧光素酶的L3.6pl细胞原位注射到无胸腺裸鼠的胰腺中，分别用（i）溶剂（1 mL/kg腹腔注射PBS，每周2次；1 mL/kg口服聚维酮/普朗尼克F68，每周3次）、（ii）KPT-330（20 mg/kg口服，每周3次）、（iii）吉西他滨（100 mg/kg腹腔注射，每周2次）或（iv）KPT-330（10 mg/kg）+吉西他滨（50 mg/kg）处理4周。与溶剂处理相比，单独使用KPT-330和吉西他滨在体外剂量依赖性地抑制贴壁依赖性生长，在体内抑制肿瘤体积。然而，联合使用具有协同抑制生长作用。联合使用时，KPT-330和吉西他滨协同作用，比单一药物治疗更能增强胰腺癌细胞死亡。机制上，KPT-330和吉西他滨促进细胞凋亡，诱导p27表达，消耗生存素，并抑制DNA修复蛋白的积累。总之，我们的数据表明，KPT-330通过抑制肿瘤生长、消耗抗凋亡蛋白和诱导细胞凋亡来增强吉西他滨对人胰腺癌的抗肿瘤活性。
