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基于PI3K/AKT信号通路探讨黄芪甲苷-聚乙二醇化丝素蛋白纳米粒对前列腺癌细胞增殖、迁移及自噬的作用机制

[Explore the mechanism of astragaloside IV-PESV on proliferation, migration, and autophagy of prostate cancer cells based on the PI3K/AKT signaling pathway].

作者信息

You Xu-Jun, Wen Zheng, Zheng Qing-Xia, Li Qi-Xin, Fu Wei, Li Hai-Song, Wang Bin

机构信息

Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China.

Bao'an District Traditional Chinese Medicine Hospital in Shenzhen, Shenzhen, Guangdong 518133, China.

出版信息

Zhonghua Nan Ke Xue. 2023 Dec;29(12):963-972.

Abstract

OBJECTIVE

Explore the effects of Astragaloside IV and Scorpion Venom Peptide on the activity, migration, apoptosis, cell cycle, autophagy, and the expression of proteins related to the PI3K/AKT signaling pathway in prostate cancer cells.

METHODS

The human prostate cancer cell lines LNCaP and PC-3 were randomly divided into blank control group, Astragaloside IV group, Scorpion Venom Peptide group, Astragaloside IV-Scorpion Venom Peptide group, and rapamycin (positive drug group). After corresponding drug treatments for 24 hours, logarithmic growth phase tumor cells were collected for testing. Cell proliferation was assessed using a Cell Counting Kit-8 (CCK-8) assay, Transwell assay, apoptosis assay, cell cycle assay, and immunofluorescence analysis were performed to detect the activity and migration capacity of prostate cancer cells in each group, as well as their effects on apoptosis, cell cycle, and the autophagy target LC3. Western blot analysis was employed to measure the protein expression levels of p-PI3K, p-Akt, p-mTOR, Beclin1, LC3, and P62.

RESULTS

Compared to the blank control group, the Astragaloside IV-Scorpion Venom Peptide group exhibited a significant decrease in the activity of prostate cancer cells (P<0.05) and a reduction in the cell invasion ability (migration capacity) (P<0.05). The early apoptosis rate (LR), late apoptosis rate (UR), and total apoptosis rate all increased (P<0.05). The proportion of cells in the G1 phase increased (P<0.05), while the proportion in the G2+S phase decreased (P<0.05). The immunofluorescence expression of LC3 significantly increased (P<0.05). The expression of LC3Ⅱ and Beclin1 proteins in prostate cancer cells LNCaP and PC-3 was upregulated (P<0.05), while the expression of P62, p-PI3K, p-AKT, and p-mTOR proteins was downregulated (P<0.05).Astragaloside IV-Scorpion Venom Peptide is superior to the Astragaloside IV group or Scorpion Venom Peptide group alone in inhibiting the activity and migration capacity of prostate cancer cells, suppressing cell mitosis, promoting early apoptosis, upregulating the expression level of LC3, and inhibiting the PI3K/AKT pathway while promoting autophagy (P<0.05).

CONCLUSION

The mechanism by which Astragaloside IV-Scorpion Venom Peptide inhibits the proliferation and migration of prostate cancer cells, suppresses cell mitosis, promotes early apoptosis, and enhances autophagy may be related to the inhibition of the PI3K/AKT pathway.

摘要

目的

探讨黄芪甲苷IV和蝎毒肽对前列腺癌细胞活性、迁移、凋亡、细胞周期、自噬以及PI3K/AKT信号通路相关蛋白表达的影响。

方法

将人前列腺癌细胞系LNCaP和PC-3随机分为空白对照组、黄芪甲苷IV组、蝎毒肽组、黄芪甲苷IV-蝎毒肽组和雷帕霉素(阳性药物组)。相应药物处理24小时后,收集对数生长期肿瘤细胞进行检测。采用细胞计数试剂盒-8(CCK-8)法评估细胞增殖,进行Transwell实验、凋亡检测、细胞周期检测以及免疫荧光分析,以检测各组前列腺癌细胞的活性和迁移能力,以及它们对凋亡、细胞周期和自噬靶点LC3的影响。采用蛋白质免疫印迹法检测p-PI3K、p-Akt、p-mTOR、Beclin1、LC3和P62的蛋白表达水平。

结果

与空白对照组相比,黄芪甲苷IV-蝎毒肽组前列腺癌细胞活性显著降低(P<0.05),细胞侵袭能力(迁移能力)下降(P<0.05)。早期凋亡率(LR)、晚期凋亡率(UR)和总凋亡率均升高(P<0.05)。G1期细胞比例增加(P<0.05),而G2+S期细胞比例下降(P<0.05)。LC3的免疫荧光表达显著增加(P<0.05)。前列腺癌细胞系LNCaP和PC-3中LC3Ⅱ和Beclin1蛋白表达上调(P<0.05),而P62、p-PI3K、p-AKT和p-mTOR蛋白表达下调(P<0.05)。黄芪甲苷IV-蝎毒肽在抑制前列腺癌细胞活性和迁移能力、抑制细胞有丝分裂、促进早期凋亡、上调LC3表达水平以及抑制PI3K/AKT通路并促进自噬方面优于单独的黄芪甲苷IV组或蝎毒肽组(P<0.05)。

结论

黄芪甲苷IV-蝎毒肽抑制前列腺癌细胞增殖和迁移、抑制细胞有丝分裂、促进早期凋亡以及增强自噬的机制可能与抑制PI3K/AKT通路有关。

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