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目的 探究Serpinc1基因通过调控PI3K/AKT信号通路对白蛋白结合型紫杉醇（nab-PTX）联合程序性死亡受体-1(PD-1)抗体抗肺癌的效果及其机制。方法 利用Serpinc1基因敲低的稳定转染的小鼠Lewis细胞系构建肺癌皮下移植瘤小鼠模型，将造模成功的150只小鼠根据实验目的随机分为NC组、NC+nab-PTX组、NC+PD-1抗体组、NC+联合治疗组、shRNA组、shRNA+nab-PTX组、shRNA+PD-1抗体组、shRNA+联合治疗组、shRNA+740 Y-P组、shRNA+nab-PTX+740 Y-P组、shRNA+PD-1抗体+740 Y-P组、shRNA+联合治疗+740 Y-P组。测量小鼠肿瘤体积和肿瘤质量，记录小鼠生存期，对肿瘤组织和肺组织进行苏木素-伊红染色，观察肿瘤转移情况。采用Western Blot法检测肿瘤组织中Serpinc1蛋白表达水平，以及PI3K、AKT和NF-κB磷酸化水平，采用免疫组织化学（IHC）法检测肿瘤组织中Ki67和TLR4的表达水平。结果 shRNA组小鼠的肿瘤体积、肿瘤质量和肺部转移灶数量低于NC组（P<0.05），小鼠生存期长于NC组（P<0.05）。与其他组比较，shRNA+联合治疗组小鼠肿瘤增长最慢（P<0.05），相对肿瘤质量最低（P<0.05）,Ki67阳性表达细胞数量最少（P<0.05）。740 Y-P组小鼠肿瘤体积增长最快（P<0.05），肿瘤组织中PI3K和AKT的磷酸化水平最高（P<0.05）;shRNA+联合治疗+740 Y-P组小鼠肿瘤体积、肿瘤质量、Ki67阳性表达细胞数量和PI3K/AKT蛋白的磷酸化水平与shRNA+nab-PTX+740 Y-P组、shRNA+PD-1抗体+740Y-P组比较，差异无统计学意义（P > 0.05）。shRNA组和shRNA+联合治疗组小鼠肿瘤组织TLR4表达水平均降低（P<0.05）；与shRNA组比较，shRNA+740 Y-P组NF-κB的磷酸化水平增加（P<0.05）,TLR4表达水平没有显著改变（P > 0.05）。结论 Serpinc1基因可能通过调节TLR4的表达水平调控PI3K/AKT信号通路，增强nab-PTX联合PD-1抗体的抗肺癌效果。
Abstract：AIM To investigate the efficacy and mechanisms of nab-PTX combined with anti-PD-1 antibody in treating lung cancer through the regulation of the PI3K/AKT signaling pathway. METHODS The mouse Lewis cells stably transfected with the Serpinc1 gene knockdown were used to construct a mouse model of subcutaneous transplanted tumor of lung cancer.The mice were randomly divided into different groups: the NC group, NC + nab-PTX group, NC + anti-PD-1 antibody group,NC + combined treatment group, shRNA group, shRNA + nab-PTX group, shRNA + anti-PD-1 antibody group, shRNA +combined treatment group, shRNA + 740 Y-P group, shRNA + nab-PTX + 740 Y-P group, shRNA + anti-PD-1 antibody + 740Y-P group, and shRNA + combined treatment + 740 Y-P group. The anti-lung cancer effects of the Serpinc1 gene and its effects on the anti-lung cancer efficacy of nab-PTX combined with anti-PD-1 antibody in vivo were investigated by measuring tumor volume, tumor weight, survival time, hematoxylin and eosin(HE) staining in tumor tissues and lung tissues. The expression of Serpinc1 protein, and the phosphorylation levels of PI3K, AKT and NF-κB in tumor tissues were investigated by Western Blot, and the expression levels of Ki67 and TLR4 in tumor tissues were detected by immunohistochemistry(IHC). RESULTS The tumor volume, tumor weight and lung metastases of mice in the shRNA group were significantly decreased than those in the NC group, and their survival time was longer than that of the NC group. Following combined treatment with nab-PTX and anti-PD-1 antibody, the shRNA group mice demonstrated the slowest tumor growth(P < 0.05), the lowest relative tumor weight(P < 0.05), and the lowest Ki67 expression level(P < 0.05) among all groups. Mice in the 740 Y-P group exhibited the most rapid tumor volume increase(P < 0.05), and the highest levels of phosphorylated PI3K and AKT in tumor tissues(P < 0.05).There were no statistically significant differences in tumor volume, tumor weight, Ki67 expression level and the phosphorylation level of PI3K/AKT protein between shRNA+ combined treatment group and shRNA+nab-PTX+740 Y-P group as well as the shRNA + anti-PD-1 antibody + 740 Y-P group(P > 0.05). Both the Serpinc1 gene knockdown and the combination treatment could inhibit the expression of TLR4 in tumor tissues, and the phosphorylation level of NF-κB was significantly increased after the administration of 740 Y-P, while the expression of TLR4 was not significantly changed.CONCLUSION The Serpinc1 gene may regulate the PI3K/AKT signaling pathway through TLR4 to enhance the efficacy of nab-PTX combined with anti-PD-1 antibody in lung cancer treatment.
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基本信息:
DOI：10.19577/j.1007-4406.2025.01.001
中图分类号:R734.2
引用信息:
[1]张君,郝文婧,王云霞,等.Serpinc1基因通过调控PI3K/AKT信号通路增强白蛋白结合型紫杉醇联合程序性死亡受体-1抗体的抗肺癌效果[J].中国临床药学杂志,2025,34(01):1-9.DOI:10.19577/j.1007-4406.2025.01.001.
基金信息:
国家自然科学基金（编号82173892）
2025-01-25
2025-01-25