目的 研究灯盏花乙素对子宫内膜癌Ishikawa细胞的影响,并分析与磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号通路的相关性.方法 将Ishikawa细胞分为空白组及低、中、高剂量实验组,分别用含0、5、10及20μmol·L-1的灯盏花乙素的完全培养基处理.用细胞计数试剂盒-8(CCK-8)实验检测细胞活力;用平板克隆实验检克隆形成能力;用划痕及Transwell实验检测转移及侵袭能力;用流式细胞术检测凋亡;用蛋白质印迹法检测蛋白表达情况.结果 空白组和低、中、高剂量实验组48 h的细胞活力分别为(100.00±0.00)％、(78.51±7.54)％、(52.93±4.91)％和(41.62±5.33)％;克隆形成率分别为(100.00±0.00)％、(56.59±6.34)％、(35.23±4.62)％和(10.66±1.91)％;划痕愈合率分别为(53.70±6.19)％、(40.59±4.75)％、(34.25±4.40)％和(15.78±2.14)％;侵袭细胞数分别为(189.70±14.06)、(106.82±12.67)、(84.37±8.13)和(53.74±6.78)个;基质金属蛋白酶-2(MMP-2)蛋白相对表达水平分别为0.96±0.10、0.73±0.06、0.68±0.08 和0.42±0.05;MMP 抑制药-1(TIMP-1)蛋白相对表达水平分别为 0.35±0.04、0.51±0.05、0.74±0.08和 1.20±0.14;细胞凋亡率分别为(4.21±0.53)％、(15.83±2.42)％、(22.72±3.85)％和(34.41±4.67)％;B 淋巴细胞瘤-2(Bcl-2)蛋白表达水平分别为 1.38±0.15、0.90±0.10、0.56±0.06 和 0.24±0.03;Bcl-2 相关 X 蛋白(Bax)表达水平分别为 0.31±0.02、0.44±0.04、0.93±0.11 和 1.26±0.14;PI3Kp85 蛋白表达水平分别为 0.67±0.05、0.42±0.04、0.36±0.02 和0.28±0.03;磷酸化 Akt(p-Akt)蛋白表达水平分别为 0.78±0.06、0.53±0.04、0.46±0.05和0.42±0.03;低、中、高剂量实验组上述指标与空白组比较,差异均有统计学意义(P＜0.05或P＜0.01).结论 灯盏花乙素可以通过调控PI3K/Akt信号通路抑制Ishikawa细胞增殖、转移及侵袭,并诱导其凋亡.
Objective To study the effects of scutellarin on endometrial carcinoma Ishikawa cells,and analyze the correlation between the effects and phosphatidyl inositol 3 kinase(PI3 K)/protein kinase B(Akt)signaling pathway.Methods Ishikawa cells were divided into blank group and experimental-L,-M,-H groups,each group was treated with complete medium containing 0,5,10 and 20 μmol·L-1 scutellarin,respectively.Cell viability,clonal formation ability,metastatic ability,invasion,apoptosis and protein expression were detected by cell counting kit-8(CCK-8),plate cloning,scratch,Transwell,flow cytometry and Western blot assay,respectively.Results The cell viability of blank group and experimental-L,-M,-H groups at 48 h were(100.00±0.00)％,(78.51±7.54)％,(52.93±4.91)％and(41.62±5.33)％;the clone formation rate were(100.00±0.00)％,(56.59±6.34)％,(35.23±4.62)％and(10.66±1.91)％;the scratch healing rate were(53.70±6.19)％,(40.59±4.75)％,(34.25±4.40)％and(15.78±2.14)％;the number of invasive cells were 189.70±14.06,106.82±12.67,84.37±8.13 and 53.74±6.78;the relative expression levels of matrixmetallo proteinase-2 were 0.96±0.10,0.73±0.06,0.68±0.08 and 0.42±0.05;tissue inhibitor of MMP-1(TIMP-1)were 0.35±0.04,0.51±0.05,0.74±0.08 and 1.20±0.14;the apoptosis rates were(4.21±0.53)％,(15.83±2.42)％,(22.72±3.85)％and(34.41±4.67)％;the relative expression levels of B cell lymphoma-2(Bcl-2)were 1.38±0.15,0.90±0.10,0.56±0.06 and 0.24±0.03;Bcl-2 associated X protein(Bax)were 0.31±0.02,0.44±0.04,0.93±0.11 and 1.26±0.14;the relative expression levels of PI3Kp85 were 0.67±0.05,0.42±0.04,0.36±0.02 and 0.28±0.03;phosphorylated Akt(p-Akt)were 0.78±0.06,0.53±0.04,0.46±0.05 and 0.42±0.03.Compared with the blank group,the above indexes in the experimental-L,-M,-H groups were statistically significant(P＜0.05 or P＜0.01).Conclusion Scutellarin can inhibit the proliferation,metastatic ability and invasion of endometrial carcinoma Ishikawa cells and promote apoptosis by regulating PI3K/Akt signaling pathway.
代丽丽;王景;魏许瑞;马庆亚;王娜;都治香
河北医科大学第一医院产科,河北石家庄 050000河北医科大学第一医院妇科,河北石家庄 050000
中医学
灯盏花乙素;子宫内膜癌;转移;侵袭;凋亡
scutellarin;endometrial carcinoma;metastatic ability;invasion;apoptosis
《中国临床药理学杂志》 2024 (001)
27-31 / 5
河北省医学科学课题研究计划基金资助项目(20210081)
10.13699/j.cnki.1001-6821.2024.01.006
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