雞屎藤苷酸誘導SGC7901胃癌細胞凋亡并抑制細胞惡性增殖和侵襲

郭強　李井野　王威　兆勇

摘要目的：初步探索雞屎藤苷酸（PA）誘導SGC7901胃癌細胞凋亡并抑制細胞惡性增殖和侵襲的可能機制。方法：選取SGC7901胃癌細胞為研究對象，利用CCK8法進行濃度梯度實驗確定后續實驗濃度，利用流式細胞術檢測0、20、40、80 μg/mL的PA處理后SGC7901胃癌細胞凋亡率以及線粒體膜電位變化情況，BrdU法和Transwell小室實驗SGC7901胃癌細胞的增殖活性以及侵襲能力，qRTPCR檢測抑癌基因P53、P21的mRNA相對表達水平，Western Blotting檢測凋亡相關蛋白Bcl2、Bax、Caspase3、cMyc和轉移相關蛋白VEGF、波形蛋白表達水平。結果：CCK8梯度濃度試驗發現PA濃度從40 μg/mL開始對SGC7901胃癌細胞活性產生了明顯抑制作用（P<0.05），后續以20、40、80 μg/mL為試驗濃度;與0 μg/mL的PA處理比較，40、80 μg/mL的PA處理后SGC7901胃癌細胞凋亡率、Bax/Bcl2、CleavedCaspase3/Caspase3蛋白表達水平以及P53、P21相對表達量升高，線粒體膜電位、細胞增殖活性、侵襲率、cMyc、VEGF、波形蛋白表達水平降低，差異均有統計學意義（均P<0.05）。結論：PA可能通過降低線粒體膜電位促進線粒體凋亡途徑誘導細胞凋亡，促進抗癌基因表達抑制細胞的惡性增殖，并抑制上皮間質轉化降低侵襲能力。

關鍵詞胃癌;SGC7901胃癌細胞;雞屎藤苷酸;惡性增殖;侵襲;轉移;凋亡;線粒體膜電位

Paederosidic Acid Induces Apoptosis of Gastric Cancer SGC7901 Cells and Inhibits

Their Malignant Proliferation and Invasion

GUO Qiang，LI Jingye，WANG Wei，ZHAO Yong

（Department of General Surgery，Benxi Central Hospital，Benxi 111027，China）

AbstractObjective：To preliminarily explore the underlying mechanism of paederosidic acid（PA） in inducing the apoptosis of gastric cancer SGC7901 cells and inhibiting their malignant proliferation and invasion.Methods：The concentration gradient test was conducted on SGC7901 cells by CCK8 to determine the subsequent experimental concentration.The changes in the apoptosis rate of SGC7901 cells and mitochondrial membrane potential（MMP） after PA treatment（0，20，40，and 80 μg/mL） were detected by flow cytometry.The proliferation activity and invasion ability of SGC7901 cells were detected by BrdU staining and Transwell assay.The relative expression levels of tumor suppressor genes（P53 and P21） were detected by qRTPCR.The expression levels of apoptosisrelated proteins（Bcl2，Bax，Caspase3，and cMyc） and metastasisrelated proteins（VEGF and vimentin） were detected by Western blot.Results：As revealed by the CCK8 concentration gradient test，PA at the concentration higher than 40 μg/mL showed inhibitory effects on the activity of SGC7901 cells（P<0.05）.The subsequent experimental concentrations were determined as 20，40，and 80 μg/mL.Compared with the results after PA treatment at 0 μg/mL，the apoptosis rate of SGC7901 cells，protein expression levels of Bax/Bcl2 and cleavedCaspase3/Caspase3，and relative mRNA expression levels of P53 and P21 increased，while MMP，cell proliferation activity，invasion rate，and protein expression levels of cMyc，VEGF，and vimentin decreased after PA treatment at 40 and 80 μg/mL（P<0.05）.Conclusion：PA may induce cell apoptosis by reducing MMP and promoting mitochondrial apoptosis pathway to promote the expression of antioncogenes，inhibit malignant proliferation and epithelialmesenchymal transition，and reduce the invasion ability.

KeywordsGastric cancer; SGC7901; Paederosidic acid; Malignant proliferation; Invasion; Metastasis; Apoptosis; Mitochondrial membrane potential

中圖分類號：R273;R735.2文獻標識碼：Adoi：10.3969/j.issn.1673-7202.2022.04.008

胃癌是一類常見的消化系統惡性腫瘤，多以腺癌為主，根據世界衛生組織統計結果顯示，亞洲范圍內胃癌的發病率占據全球惡性腫瘤發生率的第7位，死亡率在10%左右[12]，在我國胃癌的發病率僅次于第1位的肺癌，同時胃癌死亡率在所有惡性腫瘤中排在第3位[3]。化療作為晚期胃癌患者的主要治療方式之一，通常會伴隨較為嚴重的不良反應，而對于多重耐藥的GC患者而言，化療的臨床受益極低[4]，因此尋找治療胃癌的新方法具有重要的意義。……