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芥子碱硫氰酸盐对人皮肤鳞状细胞癌SCL-1细胞增殖、上皮间质转化、转移的影响及其机制研究
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| 篇名： | 芥子碱硫氰酸盐对人皮肤鳞状细胞癌SCL-1细胞增殖、上皮间质转化、转移的影响及其机制研究 |
| TITLE： | Study on the Effects and Its Mechanism of Sinapine Thiocyanate on the Proliferation ，Epithelial Mesenchymal Transition and Metastasis of Human Cutaneous Squamous Cell Carcinoma SCL- 1 Cells |
| 摘要： | 目的：研究芥子碱硫氰酸盐（ST）对人皮肤鳞状细胞癌SCL-1细胞增殖、上皮间质转化（EMT）和转移的影响，并考察其可能的作用机制。方法：将人皮肤鳞状细胞癌SCL-1细胞分为空白对照组（0.1％二甲基亚砜）和ST不同浓度组（5、10、20μmol/L），分别通过CCK-8实验、5-乙炔基-2′脱氧尿嘧啶核苷染色实验、细胞划痕实验和Transwell小室侵袭实验测定各组细胞的增殖、迁移和侵袭能力，通过Westernblot实验和免疫荧光实验分别测定各组细胞中EMT相关指标N-钙黏着蛋白（N-cadherin）、E-钙黏着蛋白（E-cadherin）的表达水平。另将SCL-1细胞分为空白对照组（0.1％二甲基亚砜）、ST单用组（20μmol/L）、ST+NSC228155组[20μmol/LST+100μmol/LNSC228155（EGFR激动剂）]和ST+SC79组[20μmol/LST+20μmol/LSC79（PI3K/Akt激动剂）]，分别通过CCK-8实验、细胞划痕实验和Transwell小室侵袭实验测定各组细胞的增殖、迁移和侵袭能力，并通过Westernblot实验测定空白对照组（0.1％二甲基亚砜）和ST不同浓度组（5、10、20μmol/L）组细胞中表皮生长因子受体（EGFR）、磷脂酰肌醇3激酶（PI3K）、磷酸化磷脂酰肌醇3激酶（p-PI3K）、蛋白激酶B（Akt）和磷酸化Akt（p-Akt）蛋白的表达水平，以验证ST的作用与EGFR/PI3K/Akt信号通路的关系。另将SCL-1细胞和人正常皮肤成纤维细胞WS1分别分为空白对照组（0.1％二甲基亚砜）、ST组（20μmol/L）、ZD1839组（阳性对照，20μmol/L，EGFR抑制剂）和LY294002组（阳性对照，20μmol/L，PI3K/Akt抑制剂），采用CCK-8实验测定各组细胞的增殖能力，以评价ST的细胞毒性。结果：与空白对照组比较，5、10、20μmol/LST组SCL-1细胞的增殖、迁移和侵袭能力均显著减弱（P＜0.05）；Westernblot和免疫荧光实验结果显示，5、10、20μmol/LST组SCL-1细胞中N-cadherin蛋白的表达均显著下调（P＜0.05），E-cadherin蛋白的表达均显著上调（P＜0.05），并且细胞中EGFR、p-PI3K、p-Akt蛋白的表达水平均显著降低（P＜0.05）。与ST单用组比较，ST+NSC228155组和ST+SC79组SCL-1细胞的增殖、迁移和侵袭能力均显著增强（P＜0.05）。与空白对照组比较，ST组WS1细胞的增殖能力差异无统计学意义（P＞0.05），SCL-1细胞增殖能力显著减弱（P＜0.05），ZD1839组和LY294002组两种细胞的增殖能力均显著减弱（P＜0.05）；与ST组比较，ZD1839组和LY294002组WS1细胞的增殖能力均显著减弱（P＜0.05），但SCL-1细胞的增殖能力差异无统计学意义（P＞0.05）。结论：ST可能通过抑制EGFR/PI3K/Akt信号通路的活化而抑制人皮肤鳞癌SCL-1细胞的增殖、EMT和转移，且其毒副作用较小。 |
| ABSTRACT： | OBJECTIVE：To stud y the effects of sinapine thiocyanate （ST） on the proliferation ，epithelial mesenchymal transformation（EMT）and metastasis of human cutaneous squamous cell carcinoma SCL- 1 cells，and to investigate its possible mechanism. METHODS ：Human cutaneous squamous cell carcinoma SCL- 1 cells were divided into blank control group （0.1％ DMSO） and ST different concentration groups （5，10，20 μmol/L）. CCK- 8 assay，5-ethynyl-2′-deoxyuridine（EDU）test， scratch test and Transwell chamber invasion test were adopted to test the proliferation ，migration and invasion ability. The expression of N-cadherin and E-cadherin were detected by Western blot and immunofluorescence assay . Other SCL- 1 cells were collected and divided into blank control group （0.1％ DMSO），ST group （20 μmol/L），ST+NSC228155 group [ 20 μmol/L ST+100 μmol/L NSC228155（EGFR agonist ）] and ST+SC 79 group [ 20 μmol/L ST+20 μmol/L SC79（PI3K/Akt agonist ）]. The proliferation ，migration and invasion ability of SCL- 1 cells in each group were detected by CCK- 8 assay，scratch test and Transwell chamber invasion assay. The expression of epidermal growth factor receptor （EGFR），phosphatidylinositol 3 kinase（PI3K），phosphorylated phosphatidylinositol 3 kinase（p-PI3k），protein kinase B （Akt）and phosphorylated protein Akt （p-Akt）protein of cells in blank control group and ST different concentration groups（5，10，20 μmol/L）were determined by Western blot assay so as to validate the relationship between ST effect and EGFR/ PI3K/Akt signaling pathway. SCL- 1 cells and human normal skin fibroblasts cell WS 1 were divided into blank control group （0.1％ DMSO），ST group （20 μmol//L），ZD1839 group（positive control ，20 μmol//L，EGFR inhibitor ）and LY 294002 group（positive control，20 μmol//L，PI3K/Akt inhibitor ）. CCK- 8 assay was used to detect the cell proliferation in order to evaluate the cells cytotoxicity of ST. RESULTS ：Compared with blank control group ，the proliferation ，migration and invasion ability of SCL- 1 cells were significantly decreased in 5，10，20 μmol/L ST groups（P＜0.05）. Western blot and immunofluorescence assay showed that the expression of N-cadherin in SCL- 1 cells were decreased significantly in 5，10，20 μmol/L ST groups（P＜0.05），while the protein expression of E-cadherin was increased significantly （P＜0.05）；the protein expressions of EGFR ，p-PI3K and p-Akt were significantly decreased （P＜0.05）. Compared with ST group ，the proliferation ，migration and invasion ability of SCL- 1 cells were increased significantly in ST + NSC 228155 group and ST + SC 79 group （P＜0.05）. Compared with blank control group ，the proliferation ability of WS 1 cells had no significant change in ST group ，while the proliferation ability of SCL- 1 cells was decreased significantly （P＜0.05）；the proliferation ability of the two kinds of cells were decreased significantly in ZD 1839 group and LY 294002 group（P＜0.05）. Compared with ST group ，the proliferation ability of WS 1 cells was decreased significantly in ZD1839 group and LY 294002 group（P＜0.05），but there was no significant difference in the proliferation ability of SCL- 1 cells （P＞0.05）. CONCLUSIONS ：ST may inhibit the proliferation ，EMT and metastasis of SCL- 1 cells through inhibiting the activation of EGFR/PI 3K/Akt signaling pathway ，and its side effects are few. |
| 期刊： | 2021年第32卷第08期 |
| 作者： | 苏羽屾，曾智锐，荣冬芸，王叶，李丹，唐姗姗，汪涛，龙雪梅，曹煜 |
| AUTHORS： | SU Yushen， ZENG Zhirui，RONG Dongyun，WANG Ye，LI Dan，TANG Shanshan，WANG Tao，LONG Xuemei，CAO Yu |
| 关键字： | 芥子碱硫氰酸盐；人皮肤鳞状细胞癌SCL-1细胞；增殖；上皮间质转化；转移；EGFR/PI3K/Akt信号通路 |
| KEYWORDS： | Sinapine thiocyanate ； Human cutaneous squamous cell carcinoma SCL-1 cells； Proliferation； Epithelial |
| 阅读数： | 244 次 |
| 本月下载数： | 5 次 |
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