Department of Obstetrics and Gynecology, Obstetrics and Gynecology Hospital of Fudan University, 128 Shenyang Road, Shanghai, 200090, China.
J Ovarian Res. 2023 Nov 8;16(1):212. doi: 10.1186/s13048-023-01285-0.
The present study aimed to investigate whether baicalein improves the sensitivity of resistant ovarian cancer cells to cisplatin.
Transcriptomic sequencing and bioinformatics analysis were used to screen differentially expressed CirSLC7A6 in A2780 and A2780/CDDP cells. RT-qPCR was performed to examine the expression levels of CirSLC7A6, miR-2682-5p, and SLC7A6. Cell proliferation and apoptosis were examined using a Cell Counting Kit-8 assay and flow cytometry, and cell migration and invasion were analyzed using wound healing and Transwell assays. Cell suspensions were inoculated into the subcutaneous tissues of the bilateral interscapular region of nude mice. Saline, cisplatin, baicalein and cisplatin plus baicalein were intraperitoneally injected to observe the effects on tumor growth. Toxicity analyses in the liver and kidney were performed using H&E staining. RT-qPCR and immunohistochemistry were used to detect the expression of CirSLC7A6, miR-2682-5p, and SLC7A6 in tumor tissues, and western blot analysis was carried out to measure protein expression levels.
CirSLC7A6 was markedly upregulated in A2780/CDDP cells compared with the A2780 cells. CirSLC7A6 knockdown notably increased the expression of miR-2682-5p and decreased SLC7A6 expression. The rates of inhibition and apoptosis in the group treated with a combination of cisplatin and baicalein were significantly higher than those of the cisplatin and baicalein groups of A2780/CDDP shCirSLC7A6 cells. In A2780/CDDP shCirSLC7A6 cells, migration and invasion were significantly higher in the cisplatin and baicalein groups, compared with the combined treatment group. In the A2780/CDDP shCirSLC7A6 cell xenograft, the tumor weight of the combined treatment group was significantly lower than that of the cisplatin and baicalein groups. In addition, the combination of cisplatin and baicalein did not induce higher levels of toxicity in the liver or kidney. Baicalein alone and in combination with cisplatin notably reduced the expression of CirSLC7A6 and SLC7A6, and increased the expression of miR-2682-5p in the A2780/CDDP shCirSLC7A6 cell xenograft. In A2780/CDDP shCirSLC7A6 cells, the expression levels of P-Akt, P-mTOR, P-Erk, Bcl-2 and MMP2 were lower in the combined treatment group than in the control group.
Treatment with baicalein improved the sensitivity of ovarian cancer cells to cisplatin and inhibited cell proliferation, metastasis and tumor growth.
本研究旨在探讨黄芩素是否能提高耐药卵巢癌细胞对顺铂的敏感性。
采用转录组测序和生物信息学分析筛选 A2780 和 A2780/CDDP 细胞中差异表达的 CirSLC7A6。通过 RT-qPCR 检测 CirSLC7A6、miR-2682-5p 和 SLC7A6 的表达水平。使用细胞计数试剂盒-8 检测和流式细胞术检测细胞增殖和凋亡，使用划痕愈合和 Transwell 检测分析细胞迁移和侵袭。将细胞悬液接种到裸鼠双侧肩胛间区的皮下组织中。腹腔注射生理盐水、顺铂、黄芩素和顺铂加黄芩素，观察对肿瘤生长的影响。通过 H&E 染色分析肝肾功能毒性。采用 RT-qPCR 和免疫组化检测肿瘤组织中 CirSLC7A6、miR-2682-5p 和 SLC7A6 的表达，采用 Western blot 分析检测蛋白表达水平。
CirSLC7A6 在 A2780/CDDP 细胞中的表达明显高于 A2780 细胞。CirSLC7A6 敲低显著增加了 miR-2682-5p 的表达，降低了 SLC7A6 的表达。与 A2780/CDDP shCirSLC7A6 细胞的顺铂和黄芩素组相比，顺铂和黄芩素联合治疗组的抑制率和凋亡率明显更高。在 A2780/CDDP shCirSLC7A6 细胞中，与联合治疗组相比，顺铂和黄芩素组的迁移和侵袭能力明显更高。在 A2780/CDDP shCirSLC7A6 细胞异种移植中，联合治疗组的肿瘤重量明显低于顺铂和黄芩素组。此外，顺铂和黄芩素联合使用并未导致肝脏或肾脏毒性增加。黄芩素单独和联合顺铂显著降低了 A2780/CDDP shCirSLC7A6 细胞异种移植中 CirSLC7A6 和 SLC7A6 的表达，增加了 miR-2682-5p 的表达。在 A2780/CDDP shCirSLC7A6 细胞中，与对照组相比，联合治疗组的 P-Akt、P-mTOR、P-Erk、Bcl-2 和 MMP2 表达水平较低。
黄芩素治疗可提高卵巢癌细胞对顺铂的敏感性，并抑制细胞增殖、转移和肿瘤生长。
