Wong Kaitlyn E, Mora Maria C, Sultana Nazneen, Moriarty Kevin P, Arenas Richard B, Yadava Nagendra, Schneider Sallie S, Tirabassi Michael V
1 Baystate Medical Center, University of Massachusetts Medical School, Springfield, MA, USA.
2 Pioneer Valley Life Sciences Institute, Springfield, MA, USA.
Tumour Biol. 2018 Jun;40(6):1010428318779515. doi: 10.1177/1010428318779515.
Outcomes of children with high grade neuroblastoma remain poor despite multi-agent chemotherapy regimens. Rhodiola crenulata extracts display anti-neoplastic properties against several cancers including breast cancer, melanoma, and glioblastoma. In this study, we evaluated the anti-neoplastic potential of Rhodiola crenulata extracts on human neuroblastoma cells. Through this work, cell viability and proliferation were evaluated following treatments with ethanol (vehicle control) or Rhodiola crenulata extract in neuroblastoma, NB-1691 or SK-N-AS cells, in vitro. HIF-1 transcriptional activity was evaluated using a dual luciferase assay. Quantitative real-time polymerase chain reaction was utilized to assess the expression of HIF-1 targets. Selected metabolic intermediates were evaluated for their ability to rescue cells from Rhodiola crenulata extract-induced death. Lactate dehydrogenase, pyruvate kinase, and pyruvate dehydrogenase activities and NAD/NADH levels were assayed in vehicle and Rhodiola crenulata extract-treated cells. The effects of Rhodiola crenulata extracts on metabolism were assessed by respirometry and metabolic phenotyping/fingerprinting. Our results revealed striking cytotoxic effects upon Rhodiola crenulata extract treatment, especially prominent in NB-1691 cells. As a greater response was observed in NB-1691 cells therefore it was used for remaining experiments. Upon Rhodiola crenulata extract treatment, HIF-1 transcriptional activity was increased. This increase in activity correlated with changes in HIF-1 targets involved in cellular metabolism. Serendipitously, we observed that addition of pyruvate protected against the cytotoxic effects of Rhodiola crenulata extracts. Therefore, we focused on the metabolic effects of Rhodiola crenulata extracts on NB-1691 cells. We observed that while the activities of pyruvate kinase and pyruvate dehydrogenase activities were increased, the activity of lactate dehydrogenase activity was decreased upon Rhodiola crenulata extract treatment. We also noted a decline in the total NAD pool following Rhodiola crenulata extract treatment. This correlated with decreased cellular respiration and suppressed utilization of carbon substrates. Through this work, we observed significant cytotoxic effects of Rhodiola crenulata extract treatment upon treatment on NB-1691 cells, a human neuroblastoma cell line with MYCN amplification. Our studies suggest that these cytotoxic effects could be secondary to metabolic effect induced by treatment with Rhodiola crenulata extract.
尽管采用了多药化疗方案，高危神经母细胞瘤患儿的预后仍然很差。大花红景天提取物对包括乳腺癌、黑色素瘤和胶质母细胞瘤在内的多种癌症具有抗肿瘤特性。在本研究中，我们评估了大花红景天提取物对人神经母细胞瘤细胞的抗肿瘤潜力。通过这项工作，在体外使用乙醇（溶剂对照）或大花红景天提取物处理神经母细胞瘤NB - 1691或SK - N - AS细胞后，评估细胞活力和增殖情况。使用双荧光素酶测定法评估HIF - 1转录活性。利用定量实时聚合酶链反应评估HIF - 1靶标的表达。评估选定的代谢中间体拯救细胞免受大花红景天提取物诱导死亡的能力。在溶剂对照和大花红景天提取物处理的细胞中测定乳酸脱氢酶、丙酮酸激酶和丙酮酸脱氢酶活性以及NAD/NADH水平。通过呼吸测定法和代谢表型分析/指纹图谱评估大花红景天提取物对代谢的影响。我们的结果显示，大花红景天提取物处理后具有显著的细胞毒性作用，在NB - 1691细胞中尤为突出。由于在NB - 1691细胞中观察到更大的反应，因此它被用于其余实验。大花红景天提取物处理后，HIF - 1转录活性增加。这种活性增加与参与细胞代谢的HIF - 1靶标的变化相关。偶然地，我们观察到添加丙酮酸可保护细胞免受大花红景天提取物的细胞毒性作用。因此，我们专注于大花红景天提取物对NB - 1691细胞的代谢影响。我们观察到，大花红景天提取物处理后，丙酮酸激酶和丙酮酸脱氢酶活性增加，而乳酸脱氢酶活性降低。我们还注意到，大花红景天提取物处理后总NAD池下降。这与细胞呼吸减少和碳底物利用受抑制相关。通过这项工作，我们观察到大花红景天提取物处理对具有MYCN扩增的人神经母细胞瘤细胞系NB - 1691细胞具有显著的细胞毒性作用。我们的研究表明，这些细胞毒性作用可能继发于大花红景天提取物处理诱导的代谢效应。
