Immunology research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Anticancer Agents Med Chem. 2019;19(15):1835-1845. doi: 10.2174/1871520619666190731155859.
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or Apo2L) is a member of the Tumor Necrosis Factor (TNF) superfamily, which stimulates apoptosis in a wide range of cancer cells through binding to Death Receptors 4 and 5 (DR4/5). Nevertheless, TRAIL has noticeable anti-cancer abilities; some cancer cells acquire resistance to TRAIL, and consequently, its potential for inducing apoptosis in target cells is strongly diminished. Acute lymphoblastic leukemia MOLT-4 cell line is one of the most resistant cells to TRAIL that developed resistance to TRAIL through different pathways. TRAIL plus kaempferol was used to eliminate the resistance of the MOLT-4 cells to TRAIL.
Firstly, IC50 for kaempferol (95μM) was determined by using the MTT assay. Secondly, the viability of the MOLT-4 cells was assayed by FACS after Annexin V/PI staining, following treatment with TRAIL (50 and 100nM) and kaempferol (95μM) alone and in combination. Finally, the expression levels of the candidate genes involved in resistance to TRAIL were assayed by real-time PCR technique.
Kaempferol plus TRAIL induced apoptosis robustly in MOLT-4 cells at 12, 24 and 48 hours after treatment. Additionally, it was found that kaempferol could inhibit the expression of c-FLIP, X-IAP, cIAP1/2, FGF-8 and VEGF-beta, and conversely augment the expression of DR4/5 in MOLT-4 cells.
It is suggested that co-treatment of MOLT-4 cells with TRAIL plus kaempferol is a practical and attractive approach to eliminate cancers' resistance to TRAIL by inhibition of the intracellular anti-apoptotic proteins, upregulation of DR4/5 and also by suppression of the VEGF-beta (VEGFB) and FGF-8 expressions.
肿瘤坏死因子相关凋亡诱导配体（TRAIL 或 Apo2L）是肿瘤坏死因子（TNF）超家族的一员，通过与死亡受体 4 和 5（DR4/5）结合，可刺激广泛的癌细胞凋亡。然而，TRAIL 具有显著的抗癌能力；一些癌细胞对 TRAIL 产生耐药性，因此其诱导靶细胞凋亡的潜力大大降低。急性淋巴细胞白血病 MOLT-4 细胞系是对 TRAIL 最耐药的细胞之一，通过不同途径对 TRAIL 产生耐药性。TRAIL 联合山奈酚用于消除 MOLT-4 细胞对 TRAIL 的耐药性。
首先，通过 MTT 测定法确定山奈酚（95μM）的 IC50。其次，通过 Annexin V/PI 染色后 FACS 分析，检测 TRAIL（50 和 100nM）和山奈酚（95μM）单独和联合处理后 MOLT-4 细胞的活力。最后，通过实时 PCR 技术检测参与 TRAIL 耐药的候选基因的表达水平。
山奈酚联合 TRAIL 在处理后 12、24 和 48 小时可强烈诱导 MOLT-4 细胞凋亡。此外，研究发现山奈酚可抑制 c-FLIP、X-IAP、cIAP1/2、FGF-8 和 VEGF-beta 的表达，同时上调 MOLT-4 细胞中 DR4/5 的表达。
研究表明，联合使用 TRAIL 和山奈酚治疗 MOLT-4 细胞是一种实用且有吸引力的方法，可通过抑制细胞内抗凋亡蛋白、上调 DR4/5 以及抑制 VEGF-beta（VEGFB）和 FGF-8 的表达来消除癌症对 TRAIL 的耐药性。
