Wang Lihua, Zhang Xuan, Wang Liangliang, Wang Beibei, Zhang Jing, Li Yuzhi
Department of Gynecology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Nov 30;40(11):1550-1556. doi: 10.12122/j.issn.1673-4254.2020.11.03.
To investigate the effect of interleukin-17A (IL-17A) on chemosensitivity of ovarian cancer cells to cisplatin (DDP) and explore the mechanism in light of autophagy regulation.
Ovarian cancer SKOV3 cells cultured were treated with different concentrations of DDP (1-20 μg/mL). MTT assay was used to observe the changes in proliferation of the treated cells and the effect of treatment with 100 ng/mL IL-17A for 24 h on DDP-induced apoptosis of SKOV3 cells. We then examined the expression of IL-17A receptor (IL-17RA) in SKOV3 cells using flow cytometry. Annexin V-FITC/PI double staining was used to detect the cell apoptosis rate, and early apoptosis of the cells was detected with JC-1 assay. A neutralizing monoclonal antibody (mAb) against IL-17RA was used to block IL-17RA. We also observed the effects of IL-17RA silencing mediated by a siRNA targeting IL-17RA (siRNA-IL-17RA) and treatment with 3-methyladenine (3-MA) for inhibiting autophagy on DDP-induced apoptosis of SKOV3 cells. The expressions of apoptosis-related proteins (Bcl-2, Bax, and cleaved caspase-3) and autophagy-related proteins (P62 and Beclin-1) in the treated cells were detected using Western blotting.
DDP increased the expression of IL-17RA in ovarian cancer SKOV3 cells. Treatment with IL-17A significantly reduced the susceptibility of SKOV3 cells to cisplatin-induced apoptosis ( < 0.05). DDP obviously augmented the expression of Beclin-1 and reduced the autophagy degradation substrate P62 protein in the cells ( < 0.05). IL-17A/IL-17RA strongly enhanced the DDPinducted autophagy of the cells ( < 0.05). Blocking autophagy with 3-MA significantly increased DDP- induced apoptosis of SKOV3 cells with IL-17RA silencing, lowered the expression of Bcl-2 and enhanced Bax expression in the cells ( < 0.05).
IL-17A/IL-17RA can decrease chemosensitivity of SKOV3 cells to DDP by upregulating DDP-induced autophagy.
探讨白细胞介素-17A（IL-17A）对卵巢癌细胞顺铂（DDP）化疗敏感性的影响，并从自噬调节角度探讨其机制。
用不同浓度的DDP（1 - 20μg/mL）处理培养的卵巢癌SKOV3细胞。采用MTT法观察处理后细胞的增殖变化以及100 ng/mL IL-17A处理24 h对DDP诱导SKOV3细胞凋亡的影响。然后用流式细胞术检测SKOV3细胞中IL-17A受体（IL-17RA）的表达。采用Annexin V-FITC/PI双染法检测细胞凋亡率，用JC-1法检测细胞早期凋亡。用抗IL-17RA的中和单克隆抗体（mAb）阻断IL-17RA。还观察了靶向IL-17RA的小干扰RNA（siRNA-IL-17RA）介导的IL-17RA沉默以及用3-甲基腺嘌呤（3-MA）抑制自噬对DDP诱导SKOV3细胞凋亡的影响。用蛋白质免疫印迹法检测处理后细胞中凋亡相关蛋白（Bcl-2、Bax和裂解的caspase-3）和自噬相关蛋白（P62和Beclin-1）的表达。
DDP增加了卵巢癌SKOV3细胞中IL-17RA的表达。IL-17A处理显著降低了SKOV3细胞对顺铂诱导凋亡的敏感性（<0.05）。DDP明显增加了细胞中Beclin-1的表达并降低了自噬降解底物P62蛋白水平（<0.05）。IL-17A/IL-17RA强烈增强了DDP诱导的细胞自噬（<0.05）。用3-MA阻断自噬显著增加了IL-17RA沉默的SKOV3细胞对DDP诱导的凋亡，降低了细胞中Bcl-2的表达并增强了Bax的表达（<0.05）。
IL-17A/IL-17RA可通过上调DDP诱导的自噬降低SKOV3细胞对DDP的化疗敏感性。
