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| SERPINA5对食管鳞癌细胞化疗敏感性的影响及机制研究* |
| The effect and mechanism of SERPINA5 on chemotherapy sensitivity of esophageal squamous cell carcinoma cells |
| 投稿时间：2024-07-10 修订日期：2024-07-10 |
| DOI： |
| 中文关键词: SERPINA5 食管鳞癌 化疗敏感性 PI3K/AKT信号通路 |
| 英文关键词: SERPINA5 Esophageal squamous cell carcinoma Chemotherapy sensitivity PI3K/AKT signaling pathway |
| 基金项目:省部共建中亚高发病成因与防治国家重点实验室开放课题(NoSKL-HIDCA-2020-3),自治区重点研发计划项目(课题)(No: 2020B03003、2020B03003-3) |
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| 中文摘要: |
| 目的:探索SERPINA5对食管鳞癌化疗敏感性的影响及可能机制。方法:采用Western blot检测SERPINA5在人食管黏膜上皮细胞系SHEE及食管鳞癌细胞系KYSE150、TE-1、ECA109的表达水平。选取KYSE150/TE-1细胞系转染过表达对照(oe-NC)、过表达SERPINA5 (oe-SERPINA5)慢病毒,采用CCK-8检测不同浓度(0、20、40、60、80、100μmol/L)顺铂(CDDP)处理24h的半数抑制浓度(IC50)。采用Edu和平板克隆实验检测细胞增殖,流式细胞术检测细胞凋亡。15只雌性裸鼠随机分为oe-NC组、oe-NC+CDDP组和oe-SERPINA5+CDDP组,通过皮下接种KYSE150慢病毒稳转株细胞构建裸鼠荷瘤模型,予以腹腔注射生理盐水或顺铂溶液(1次/3d),并测量各组小鼠瘤体体积及质量,进一步通过Ki67、TUNEL染色检测细胞增殖及凋亡水平。Western blot检测PI3K/AKT信号通路蛋白表达水平。结果: SERPINA5在食管鳞癌细胞中呈低表达。CCK-8实验结果表明,与oe-NC组比较,oe-SERPINA5组IC50明显降低(P<0.05)；Edu和平板克隆实验结果表明,与oe-NC+CDDP组比较,oe-SERPINA5+CDDP组细胞增殖水平降低(P<0.05)；流式细胞术结果表明,与oe-NC+CDDP组比较,oe-SERPINA5+CDDP组细胞凋亡比例增加(P<0.05)。体内动物实验结果表明,SERPINA5过表达显著抑制顺铂处理下的瘤体生长、降低肿瘤细胞增殖能力,并促进其凋亡(P<0.05)。Western blot结果表明,与oe-NC+CDDP组比较,oe-SERPINA5+CDDP组抑制 PI3K和AKT蛋白磷酸化(P<0.05)。结论:SERPINA5过表达降低食管鳞癌细胞增殖能力、诱导细胞凋亡,提高其化疗敏感性,其机制可能与抑制PI3K/AKT信号通路相关。 |
| 英文摘要: |
| Objective:To explore the effect and possible mechanism of SERPINA5 on chemotherapy sensitivity in esophageal squamous cell carcinoma. Methods: Western blot assay was conducted to detect the expression levels of SERPINA5 in human esophageal mucosal epithelial cell line SHEE and esophageal squamous cell carcinoma cell line(KYSE150, TE-1, and ECA109). KYSE150/TE-1 cells were transfected with overexpressing control (oe-NC) and overexpressing SERPINA5 (oe-SERPINA5) lentivirus. The half-maximal inhibitory concentration (IC50) of cisplatin (CDDP) at different concentrations (0, 20, 40, 60, 80, 100μmol/L) for 24 hours was detected by CCK-8 assay.Edu and plate cloning experiments were used to detect cell proliferation, and flow cytometry was used to detect cell apoptosis. Fifteen female nude mice were randomly divided into oe-NC group, oe-NC+CDDP group, and oe-SERPINA5+CDDP group. The subcutaneous tumor model in nude mouse was constructed by subcutaneous inoculation the transfected KYSE150 cells, normal saline or cisplatin solution is intraperitoneally injected (once/3d). The tumor volume and mass of each group were measured,the levels of cell proliferation and apoptosis were further detected by Ki67 and TUNEL staining. Western blot was used to detect proteins expression of PI3K/AKT signaling pathway. Results: SERPINA5 is downregulated in esophageal squamous cell carcinoma cells.CCK-8 experiment showed that the IC50 to CDDP of oe-SERPINA5 group was significantly diminished compared with the oe-NC group(P <0.05).Edu and plate cloning experiments showed that oe-SERPINA5+CDDP group had a lower level of cell proliferation compared with oe-NC+CDDP group(P<0.05); Flow cytometry showed that oe-SERPINA5+CDDP group had an increased proportion of cell apoptosis compared with oe-NC+CDDP group (P<0.05). In vivo animal experiment results showed that overexpression of SERPINA5 significantly inhibited tumor growth under cisplatin treatment, reduced tumor cell proliferation ability, and promoted apoptosis (P<0.05). Western blot results showed that oe-SERPINA5+CDDP group inhibited phosphorylation of PI3K and AKT protein compared with oe-NC+CDDP group(P<0.05). Conclusion: Overexpression of SERPINA5 reduces the proliferation ability of esophageal squamous cell carcinoma cells, induces cell apoptosis, and enhances chemotherapy sensitivity. Its mechanism may be related to the inhibition of the PI3K/AKT signaling pathway. |
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