亞硒酸鈉與黃連素對人類肺腺癌A549細胞增生、移動和轉移之影響

Effects of sodium selenite and berberine on proliferation, migration and metastasis in human lung adenocarcinoma A549 cells

吳柔潔(Rou-Jie Wu)

指導教授 ： 詹文雄

中原大學/理學院/生物科技研究所/碩士(2021年)

https://doi.org/10.6840/cycu202100665

摘要

根據衛生福利部108年的資料統計國人第一大癌症死因為肺癌，主要原因為缺乏早期診斷與治療腫瘤轉移的方法，且肺癌患者預後情形較差，而目前治療肺癌的方式是藉由手術切除腫瘤後進行化學治療及放射線治療達到殺死癌細胞，但是在進行這些治療的過程中常會殺死正常細胞，所以開發出安全且有效的抗癌藥物是目前最迫切的需求。在最近的研究中發現，黃連素（Berberine, BER）及亞硒酸鈉（Sodium selenite, SS）都具有抗氧化、抗發炎、誘導細胞凋亡以及抑制腫瘤細胞移動和侵襲能力，且硒為人類必需的微量營養素，缺乏可能增加癌症風險，因此希望能藉由兩種天然化合物，探討是否具有協同作用，達到抑制肺癌細胞增生及移動，以降低目前臨床治療藥物對於正常細胞的傷害和產生的副作用。本研究利用人類肺腺癌細胞株（A549），給予不同濃度的黃連素（5, 10, 20, 40 μM）和/或是亞硒酸鈉（5, 10, 20, 40 μM），利用細胞生長試驗、自由基測定、聚落形成、細胞移動與侵襲試驗及西方墨點法等方式，分析對肺腺癌細胞的生長、移動和侵襲能力以及凋亡的影響。結果顯示，單獨給予黃連素可顯著抑制肺腺癌細胞的生長、移動和侵襲能力；但單獨給予亞硒酸鈉只能在短時間抑制肺腺癌細胞的生長、移動和侵襲能力，而對於兩者合併使用雖然可抑制肺腺癌細胞生長、移動和侵襲能力但並未有加成效果。雖然黃連素與亞硒酸鈉單獨或是合併處理都能增加活性氧族群（Reactive oxygen species, ROS）的產生，但是在凋亡傳遞路徑的研究，發現未能活化凋亡蛋白酶-3（Caspase-3），因此後續實驗將進一步探討黃連素與亞硒酸鈉合併使用對於抑制肺腺癌生長、移動和侵襲及造成細胞死亡的調控機制，希冀能開發出安全且有效的抗癌藥物，減少肺腺癌細胞的生長及轉移能力。

關鍵字

肺腺癌 ； 黃連素 ； 亞硒酸鈉 ； 轉移

並列摘要

In 2019, the Ministry of Health and Welfare reported that lung cancer was the leading cause of cancer deaths in Taiwan. The reason for this is the limited available treatment for tumor metastasis, poor prognosis for lung cancer patients, and late diagnosis. The current method of treating lung cancer is to remove the tumor by surgery, and then perform chemotherapy and radiotherapy to kill the cancer cells. However, during these treatments, normal cells are often killed. Therefore, the development of safe and effective anti-cancer drugs is urgently needed. Recent studies have found that both berberine (BER) and sodium selenite (SS) have antioxidant, anti-inflammatory, pro-apoptotic, and tumor cell migration inhibitor effects. Furthermore, insufficient selenium, an essential micronutrient in the human body is reported to increase the risk of developing cancer. The goal of this research is to investigate whether berberine and selenium have a synergistic effect to inhibit the proliferation and migration of lung cancer cells as well as protective effects on the normal cells against the current available medication. In this study, human lung adenocarcinoma cell line (A549) was exposed to different concentrations of berberine (5, 10, 20, 40 μM) and/or sodium selenite (5, 10, 20, 40 μM). Cell viability analysis, reactive oxygen species staining, colony formation, cell migration, invasion test, and western-blot analysis were used to analyze the possible effects of berberine and sodium selenite on lung adenocarcinoma cell growth, migration, invasion ability, and apoptosis. The results show that berberine alone can significantly inhibit the growth, migration, and invasion of lung adenocarcinoma cells; however, sodium selenite can only inhibit the growth, migration, and invasion of lung adenocarcinoma cells for a short period. A549 cell line exposed to both berberine and sodium selenite causes inhibition of growth, migration, and invasion with no Additive effects. Although berberine and sodium selenite alone or in combination can increase the production of reactive oxygen species (ROS), the cell apoptosis pathway fails to activate Caspase-3. In the future, researchers need to further explore the mechanisms of the combined application of berberine and sodium selenite to inhibit the growth of lung adenocarcinoma. It is hoped that safe and effective anticancer drugs can be developed to reduce growth and metastatic ability of lung adenocarcinoma cells.

並列關鍵字

Lung adenocarcinoma ； Berberine ； Sodium selenite ； Metastasis

參考文獻

Barzegar, E., Fouladdel, S., Movahhed, T.K., Atashpour, S., Ghahremani, M.H., Ostad, S.N., Azizi, E., 2015. Effects of berberine on proliferation, cell cycle distribution and apoptosis of human breast cancer T47D and MCF7 cell lines. Iran J Basic Med Sci 18, 334-342.

Behne, D., Kyriakopoulos, A., 2001. Mammalian selenium-containing proteins. Annu Rev Nutr 21, 453-473.

Bhattacharyya, A., Chattopadhyay, R., Mitra, S., Crowe, S.E., 2014. Oxidative stress: an essential factor in the pathogenesis of gastrointestinal mucosal diseases. Physiol Rev 94, 329-354.

Brune, B., 2003. Nitric oxide: NO apoptosis or turning it ON? Cell Death Differ 10, 864-869.

Cain, K., Bratton, S.B., Cohen, G.M., 2002. The Apaf-1 apoptosome: a large caspase-activating complex. Biochimie 84, 203-214.