新穎白芷素衍生物藉由重置癌細胞代謝有效抑制人類鱗狀癌細胞之研究

白芷素為一種具有多種生物活性的中草藥，不僅可以抗癲癇痙攣或是做鎮靜劑，並且在近期研究中發現具有抗癌的效果，我們團隊2012年合成一系列白芷素的衍生物，在初步的研究當中，化合物 8苯甲酰基-4-甲基-9-苯基-2H-呋喃並[2,3-H]色烯-2-酮 (BPR2P0001S0)，對於許多不同癌症類型有顯著的抑制效果，但是目前抗癌的機制仍然還沒有完全釐清。因此，在本研究中，我們首先探討該化合物對於不同癌症類型的治療效果，結果發現，該化合物在人類鱗狀細胞癌 KB 及 Ca-922-der1有非常顯著的生長抑制效果，IC50可以達到奈米數量級，但對於AsPC1與 MDA-MB-231等腺癌細胞相對來說較不敏感，此外，對正常的人臍靜脈內皮細胞以及纖維母細胞並沒有明顯的細胞生長抑制現象。進一步以KB細胞為研究模型，在細胞群落形成的實驗中，我們發現在BPR2P0001S0治療後，將顯著的減少腫瘤細胞的增殖能力，但沒有造成細胞的死亡。進一步的研究顯示，BPR2P0001S0 會誘導細胞DNA損傷、細胞老化以及細胞週期停滯於S期。除此之外，此化合物還會造成細胞醣類代謝的途徑改變；致癌基因c-Myc以及醣解作用的酵素，六碳糖激酶2和乳酸脫氫酶A，皆受到藥物的作用而使得RNA與蛋白質的表現量皆下降。不僅如此，我們發現外加丙酮酸或乳酸，可以回復BPR2P0001S0誘發的細胞生長抑制作用。臨床前動物實驗的分析結果顯示，以靜脈注射的方式，將BPR2P0001S0投予有人類KB異種移植腫瘤的NOD/SCID小鼠，具有腫瘤抑制的效果。根據以上研究，我們發現BPR2P0001S0對鱗狀細胞癌具有細胞生長抑制、可誘發DNA 損傷與促使細胞週期停滯在S期、並具有重置癌症代謝的能力。接下來，我們將進一步證明BPR2P0001S0的治療，在上述各個現象間，彼此的調控網絡與關係。總而言之，我們認為BPR2P0001S0對人體鱗狀細胞癌具有治療潛力。

關鍵字

白芷素；抗癌藥物；癌症治療；鱗狀細胞癌；細胞停滯； DNA損傷； S期停滯；癌症代謝；瓦氏效應

並列摘要

Angelicin, which is categorized into the class of angular furanocoumarin, has been indicated as a potential compound that has multiple activities in various pharmacological substances, including anticonvulsants, intercalating agents, and tranquilizing agents. Recently, a series of novel angelicin derivatives have been synthesized by our drug discovery team. Preliminary screens indicated that many of them have potent anti-tumor proliferative effect. Among them, 8-Benzoyl-4-methyl-9-phenyl- 2H-furo[2,3-h]chromen-2-one (BPR2P0001S0) is the most potent anti-cancer agent against various human cancer cells growth. However, the mechanism of action underlying this anti-cancer effect is not fully understood. In the present study, we demonstrated that BPR2P0001S0 exerted potent anti-proliferation activity against several squamous cell carcinoma cell lines, such as KB and Ca-9-22, with IC50 values in the nanomolar range. Nevertheless, this compound was less potent on the inhibition of adenocarcinoma cell lines. No growth inhibitory effect was observed in normal cells, such as human umbilical vein endothelial cells (HUVEC) and fibroblast Detroit551. In the present study, we chose KB cell as the experimental model for further research. Clonogenic assay supported that BPR2P0001S0 treatment significantly reduce the proliferative capacity of KB cells, but did not cause cell death. Further studies showed that BPR2P0001S0 was able to induce DNA damage, senescence and cell cycle arrest in S phase. In addition, this compound could also cause changes in glucose metabolism by down-regulation of the expression level of c-Myc, hexokinase II and lactate dehydrogenase A mRNA and protein in KB cells. Moreover, cell growth inhibition induced by BPR2P0001S0 could be rescued by addition of lactate and methyl pyruvate in KB cells. Preclinical animal study demonstrated that BPR2P0001S0 exhibited anti-tumor activity against the growth of KB xenograft tumor in NOD/SCID mice. Overall, based on our results, we concluded that BPR2P0001S0 causes cancer growth inhibition, DNA damage, S phase arrest, cell senescence and cancer metabolism alteration. Although network that connects with each effect is under investigation, we can deduce that BPR2P0001S0 has potential to be a new therapeutic agent toward human squamous cell carcinoma growth.

並列關鍵字

angelicin ； anti-cancer drug ； cancer therapy ； squamous cell carcinoma ； cytostasis ； DNA damage ； S-phase arrest ； cancer metabolism ； Warburg effect

參考文獻

14 Basu, A. & Krishnamurthy, S. Cellular responses to Cisplatin-induced DNA damage. Journal of nucleic acids 2010, doi:10.4061/2010/201367 (2010).

1 Berman, J. J. Tumor taxonomy for the developmental lineage classification of neoplasms. Bmc Cancer 4, doi:Artn 88

Doi 10.1186/1471-2407-4-88 (2004).

2 Elledge, S. J. Cell cycle checkpoints: preventing an identity crisis. Science 274, 1664-1672 (1996).

3 Hartwell, L. H. & Weinert, T. A. Checkpoints: controls that ensure the order of cell cycle events. Science 246, 629-634 (1989).

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新穎白芷素衍生物藉由重置癌細胞代謝有效抑制人類鱗狀癌細胞之研究

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