青藤鹼是從防己科落葉纏繞藤本植物青藤及毛青藤的乾燥藤莖中提取一種生物鹼，近期的研究發現青藤鹼可藉由抑制腫瘤增生及誘導細胞凋亡而有抗乳癌和大腸癌的作用，進而抑制細胞遷移和侵襲之能力，但是其對抑制人類肺癌細胞轉移的效果及機轉尚不清楚。根據過去文獻指出，MMP-2和MMP-9會參與腫瘤細胞轉移及血管新生，包括肺癌皆表現大量的MMP相關酵素，因此本研究擬以體外試驗分析青藤鹼對人類肺腺細胞A549侵入作用的機制，以MTT分析不同濃度的青藤鹼對A549細胞生長的抑制作用，再進行In vitro Wound healing assay、Boyden chamber invasion assay來觀察青藤鹼抑制細胞爬行、侵入作用， 並以定量即時PCR測試青藤鹼對細胞所表現參與侵入作用的相關基因之表現。實驗數據顯示，濃度1 mM以上的青藤鹼可顯著抑制A549細胞生長，並發現青藤鹼可抑制MMP-9基因的mRNA表現，同時促進金屬蛋白?組織抑制蛋白TIMP-2及RECK基因的mRNA表現，這些研究成果顯示青藤鹼抑制肺腺癌A549細胞的爬行與侵入作用，可能具有抑制肺癌細胞轉移的潛能，將有助於進一步研究青藤鹼是否能有效抑制肺癌細胞增生與轉移的作用機轉。 Sinomenine is an isoquinoline named after its extraction from the Chinese medicinal plant Sinomenium acutum. In recent years, many reports have suggested that sinomenine has an anti-cancer function in breast and colon cancers due to its role in cell proliferation suppression and apoptosis promotion. Furthermore, sinomenine was found to reduce invasion and migration ability of several types of cells. But, the inhibitory effect of sinomenine on human lung cancer cell remains unclear. Based on past literature, MMP-2 and MMP-9 have been implicated in the pathological processes that contribute to tumor progression, such as tumor cell invasion and angiogenesis. In the present study, we examined the inhibitory effect of sinomenine on metastasis of lung cancer cell A549 in vitro. Cell viability, migration and invasion were determined by MTT method, in vitro wound healing assay and Boyden chamber invasion assay, respectively. The mRNA level of matrix metalloproteinase-2 (MMP-2), MMP-9, reversion-inducing cysteine-rich protein with kazal motifs (RECK), and tissue inhibitor of metalloproteinase-1/2 (TIMP-1/2) were measured by quantitative real-time PCR. Results demonstrate that doses more than 1 mM of Sinomenine can significantly inhibit the growth of A549 cells. Sinomenine inhibit mRNA expression of MMP-9, while promoting expression of TIMP-2 and RECK. These results suggest an attractive therapeutic potential of sinomenine for suppressing invasion of lung cancer cell.
