一种老年肺癌细胞微管变构多肽(SAP)的体外抗肺癌活性验证

Antilung cancer activity research of a microtubule structure-altering polypeptide in elderly lung cancer cells in vitro

中文关键词: 老年  多肽  肿瘤  微管结构  增殖能力  迁移能力

英文关键词: elderly  polypeptide  tumor  microtubule structure  proliferation ability  migration ability

中文摘要:

      目的 从设计合成的变构系列多肽(structure-altering polypeptide,SAP)中筛选出最佳多肽XA5,探究其可能对肺癌细胞活性的影响,为后续对老年耐受性差的肺癌患者提供基础.方法 挑选以上系列的XA5多肽和腺癌细胞系A549进行体外实验验证.CCK-8验证其对肺癌细胞活性影响,Transwell实验和划痕实验验证其对侵袭性转移能力的影响,微管聚合实验检测微管蛋白聚合判断优选多肽是否阻止微管聚合等特异性抗微管效能验证.结果 经细胞直接计数实验证实,3组不同剂量组的XA5细胞干预后细胞数量均较对照组下降(P＜0.05),CCK-8实验证实添加XA5多肽后,细胞活力较对照组明显下降(P＜0.05).Transwell实验中,每组XA5细胞迁移细胞数较对照组明显下降(P＜0.001),验证其对肺腺癌细胞侵袭有明显抑制能力.因其中2组剂量组(20 μg/mL、40 μg/mL)的划痕距离较对照组延长(P＜0.001),明确了 XA5多肽抑制肺癌细胞转移的能力,以上抑癌性能均与剂量呈正比.微管聚合检测发现XA5多肽可以明显抑制微管蛋白聚合.结论 设计合成的SAP系列多肽XA5经体外实验验证其具有高效抑制肺癌细胞数量、抑制其细胞的迁移和入侵能力并抑制微管聚合,是一款在老年耐受差肺癌患者中具备广阔开发前景的抗肺癌药物.

英文摘要:

      Objective To screen the best peptide XA5 from the synthesized structure-altering polypeptide(SAP),and explore its possible effect on the activity of lung cancer cells,so as to provide a basis for subsequent treatment of elderly lung cancer patients with poor tolerance.Methods The above series of XA5 polypeptides and adenocarcinoma cell line A549 were selected for verification in vitro.Cell counting kit-8(CCK-8)experiment was used to verify their effects on lung cancer cell activity.Transwell assay and scratch test were used to verify their effects on invasion and metastasis.Microtubule polymeriza-tion assay was used to detect microtubulin polymerization and determine whether the preferred peptides prevented microtubule polymerization.Results The direct cell counting experiment confirmed that the number of cells in the 3 groups with different doses of XA5 was lower than that in the control group(P＜0.05).CCK-8 experiment confirmed that the cell viability de-creased significantly after adding XA5 polypeptide(P＜0.05).In the Transwell experiment,the number of migrating XA5 cells in each group was significantly lower than that in the control group(P＜0.001),which confirmed its significant inhibitory effect on the invasion of lung adenocarcinoma cells.Two dose groups(20 μg/mL,40 μg/mL)had longer scratch distances than the control group(P＜0.001),which confirmed the ability of XA5 peptide to inhibit the metastasis of lung cancer cells.The above tumor inhibitory effects were proportional to the dose.Microtubule polymerization assay showed that XA5 peptide could significantly inhibit microtubule polymerization.Conclusion In vitro experiments confirms that the designed and synthe-sized SAP series polypeptide XA5 can effectively inhibit the number of lung cancer cells,inhibit their migration and invasion,and inhibit microtubule polymerization.It is a promising anti-lung cancer drug with broad development prospects in elderly lung cancer patients with poor tolerance.