一個可造成肺癌細胞CL1-5凋亡並抑制其轉移能力的雙功能抑制劑

A dual-functional inhibitor causes apoptosis and inhibits metastasis of lung cancer cell line CL1-5

蔡有緯(You-Wei Tsai)

指導教授 ： 梁博煌

國立臺灣大學/生命科學院/生化科學研究所/碩士(2019年)

https://doi.org/10.6342/NTU201902369

摘要

肺癌是致死率高且容易轉移(metastasis)到其他器官的一種癌症。研究使用的細胞株為CL1-5，為非小細胞肺癌(non-small cell lung cancer (NSCLC))，此類型的肺癌細胞為最常見的一種。根據先前研究發現，藉由I-Trp阻斷CCT-β與β-tubulin之間的交互作用可殺死癌細胞，而本研究使用的小分子－845，是NHRI合成出來的一系列殺菌化合物中經實驗篩選出最有潛力殺死CL1-5，且其結構是由之前使用電腦模擬出可逆型CCT-β與β-tubulin間交互作用之抑制劑中的基團與I-Trp之側鏈所組成。經實驗發現，845可誘導肺癌細胞發生內質網壓力(ER stress)，進而促使肺癌細胞內的鈣離子釋放至細胞質並導致細胞凋亡。透過wound healing assay、transwell assay，發現845能抑制肺癌細胞遷移(migration)及侵入(invasion)，且經由西方墨點法可知845能抑制EMT、AKT/β-catenin和integrin的訊息傳導路徑，進而抑制肺癌轉移。
總括而論，本研究發現此具有雙功能的抑制劑化合物-845具有對肺癌的抗腫瘤和抗轉移能力。

關鍵字

肺癌 ； 抗腫瘤 ； 抗轉移

並列摘要

Lung cancer has a high mortality rate and often metastasizes to other organs. The cell line used in the study was CL1-5, which is a non-small cell lung cancer (NSCLC). According to previous studies, cancer cells can be killed through blocking the interaction between CCT-β and β-tubulin by I-Trp. The small molecule compound-845 used in this study was screened among a series of bactericidal compounds synthesized by NHRI and is the most potential to kill CL1-5 cells. In addition, its structure is composed of a functional group which is similar to the reversible inhibitors of CCT-β: β-tubulin complex previously simulated by computer and a side chain of I-Trp. It has been found that compound-845 would induce ER stress in CL1-5 cells, which in turn leads to the release of intracellular calcium ion and apoptosis. Furthermore, analyzed by wound healing assay and transwell assay, it has been found that compound-845 would inhibit the migratory and invasive ability of CL1-5 cells. Moreover, compound-845 would inhibit EMT, AKT/β-catenin and integrin signaling pathway investigated by western blot, thereby inhibiting CL1-5 cell metastasis.
In conclusion, this study reveals that this dual-functional inhibitor compound-845 probably has anti-tumor and anti-metastasis ability against human lung cancer.

並列關鍵字

lung cancer ； anti-tumor ； anti-metastasis ； EMT ； AKT/β-catenin ； integrin

參考文獻

1. Rivenzon-Segal, D., et al., Sequential ATP-induced allosteric transitions of the cytoplasmic chaperonin containing TCP-1 revealed by EM analysis. Nat Struct Mol Biol, 2005. 12(3): p. 233-7.

2. Grantham, J., K.I. Brackley, and K.R. Willison, Substantial CCT activity is required for cell cycle progression and cytoskeletal organization in mammalian cells. Exp Cell Res, 2006. 312(12): p. 2309-24.

3. Dekker, C., et al., The interaction network of the chaperonin CCT. EMBO J, 2008. 27(13): p. 1827-39.

4. Brackley, K.I. and J. Grantham, Activities of the chaperonin containing TCP-1 (CCT): implications for cell cycle progression and cytoskeletal organisation. Cell Stress Chaperones, 2009. 14(1): p. 23-31.

5. Marco, S., J. L. Carrascosa, and J. M. Valpuesta, Reversible interaction of beta-actin along the channel of the TCP-1 cytoplasmic chaperonin. Biophysical J, 1994. 67(1): p. 364-8.