黃連素透過NF-κB調控基質金屬蛋白酶轉錄活性之研究

The study of berberine on transcriptionally regulation of MMP-2 expression via the down-regulation of NF-κB

賴清美(Ching-Mei Lai)

指導教授 ： 陳霈霓 ； 張雲菁

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2014年)

摘要

在國人女性十大癌症死因中，子宮頸癌為第七名，10年來標準化死亡率在女性方面以子宮頸癌降幅最大，應該歸功於國健局自民國八十四年提供子宮頸抹片篩檢後，使死亡率約下降六成。但子宮頸癌仍為女性生殖泌尿道癌症第一名。黃連素(berberine)，又稱為小蘗鹼，是一種生物鹼(alkaloid)，在中國醫學藥用歷史悠久，具有抗發炎和抗癌的生物效果。但對抑制侵襲及轉移作用方面，其詳細機轉的研究還很少。所以我們選擇用黃連素處理子宮頸癌細胞，以MTT assays及Boyden chamber來觀察，發現黃連素具有抑制子宮頸癌細胞移動和侵入的能力，但不具有細胞毒性。接著在gelatin zymography與casein zymography assay及Western blot中也發現到黃連素可以抑制子宮頸癌細胞SiHa的MMP-2及u-PA的表現，因此我們再以luciferase assay分析黃連素對MMP-2及u-PA promoter的調控，發現同樣也有抑制的結果。深入探討其作用機轉，結果顯示黃連素可顯著抑制NF-κB在細胞核中的蛋白表現，進而抑制NF-κB與DNA的結合能力，最後藉由加入NF-κB抑制劑，觀察到黃連素會使MMP-2的蛋白表現量減少。綜合上述結果得知，黃連素可能是透過抑制NF-κB來調控轉錄因子的活性，抑制癌細胞中MMP-2的表現，進而達到抑制癌細胞轉移侵襲的能力。

關鍵字

黃連素 ； 侵襲 ； 基質金屬蛋白酶-2 ； 尿激酶型纖維蛋白溶解酶原活化劑 ； NF-κB ； SiHa細胞

並列摘要

Chinese women in the top ten causes of death in cancer, cervical cancer is seventh, 10 years standardized mortality rates of cervical cancer in females with the largest decline, should be attributed to the Republican National Health Board since 1995, providing Pap smear screening after the inspection, so that the mortality rate declined by about Liu Cheng. However, cervical cancer remains the first female reproductive tract cancers. Berberine is a plant alkaloid with a long history of medicinal use in Chinese medicine. The berberine alkaloid can be found in the roots, rhizomes, and stem bark of the plants.Berberine extracts and decoctions have demonstrated significant antimicrobial activity and has several anti-inflammation and anti-cancer biological effects. However, its mechanism of action on the cell migration and invasion of human cervical cancer cells is not fully understood. Human cervical cancer cells were treated with berberine, subjected to invasion assay, motility assay and MTT assay, significantly inhibits invasive, motility in human cervical cancer cells. Zymography and promoter luciferase analysis revealed that berberine inhibits the proteinase and transcription activities of MMP-2 and u-PA, respectively.We also use NF-κB inhibitor to find that berberine inhibited NF-κB which then led to the inhibition MMP-2 expression. Overall, berberine on transcriptionally regulation of MMP-2 expression via the down-regulation of NF-κB followed by the inhibition of migration and invasion in human cervical cancer.

並列關鍵字

Berberine ； invasion ； MMP-2 ； u-PA ； NF-κB ； SiHa cells

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