1.550004 贵阳,贵州医科大学基础医学院免疫学教研室
2.550004 贵阳,贵州医科大学组织工程与干细胞实验中心
3.230031 合肥,安徽医科大学临床医学院基础医学部
Received:10 June 2022,
Revised:2022-08-12,
Published:15 December 2022
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Fang LIU, Qin ZHONG, Xian WANG, et al. Anti-melanoma SK-Mel-103 cells effect of a natural small molecule, HEP-14[J]. Journal of Army Medical University, 2022, 44(23): 2392-2400.
Fang LIU, Qin ZHONG, Xian WANG, et al. Anti-melanoma SK-Mel-103 cells effect of a natural small molecule, HEP-14[J]. Journal of Army Medical University, 2022, 44(23): 2392-2400. DOI: 10.16016/j.2097-0927.202206064.
目的
2
初步探讨天然小分子HEP-14对黑色素瘤SK-Mel-103细胞的抑制作用及相关机制。
方法
2
用不同浓度的HEP-14(2.5~40 μmol/L)处理黑色素瘤SK-Mel-103细胞后,CCK-8和平板克隆形成实验检测细胞的存活率和克隆的形成,流式细胞术检测细胞周期变化和细胞凋亡,Transwell实验检测细胞迁移能力,RT-qPCR检测CD271和STAT3基因水平的表达,Western blot检测CD271、STAT3、p-STAT3蛋白水平的表达,细胞免疫荧光检测进一步证实CD271在细胞内的表达。
结果
2
与对照组比较,天然小分子HEP-14能有效抑制SK-Mel-103细胞的生长和克隆的形成(
P
<
0.05),诱导细胞G
2
/M期阻滞(
P
<
0.05),促进细胞凋亡(
P
<
0.05)和抑制细胞的迁移。HEP-14显著抑制SK-Mel-103、SK-Mel-147和SK-Mel-28细胞STAT3的磷酸化(
P
<
0.05),下调CD271的表达(
P
<
0.05)。
结论
2
天然小分子HEP-14能有效抑制黑色素瘤细胞生长并诱导其凋亡;可能通过抑制STAT3的激活下调CD271的表达,从而抑制黑色素瘤细胞的迁移。
Objective
2
To investigate the inhibitory effect of a natural small molecule
HEP-14
on melanoma SK-Mel-103 cells and its underlying mechanism.
Methods
2
SK-Mel-103 cells were treated with HEP-14 at different concentrations (2.5~40 μmol/L). The untreated cells were used as a control. Cell viability and colony formation were respectively assessed using CCK-8 assay and colony formation assay. Flow cytometry was applied to examine cell cycle and apoptosis. Transwell assay was used to evaluate cellular mobility. Real-time RT-qPCR was used to measure the mRNA expression of CD271 and STAT3
and Western blotting was employed to detect the protein levels of CD271
STAT3 and p-STAT3. Immunofluorescence assay was perform to confirm the expression of CD271.
Results
2
HEP-14 significantly inhibited the growth and colony formation of SK-Mel-103 cells (
P
<
0.05)
induced cell arrest at G2/M phase (
P
<
0.05)
promoted cell apoptosis (
P
<
0.05) and suppressed cell mobility in comparison with the untreated cells. In addition
HEP-14 treatment suppressed the phosphorylation of STAT3 (
P
<
0.05) and down-regulated the expression of CD271 (
P
<
0.05) in the melanoma cell lines SK-Mel-103
SK-Mel-147 and SK-Mel-28.
Conclusion
2
Natural small molecule HEP-14 effectively inhibits the proliferation and induces the apoptosis of melanoma SK-Mel-103 cells
and it suppresses the cell migration by inhibiting activation of STAT to down-regulate CD271 expression.
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