Tian Y P, Cui Y S, Zheng X, Liu B L, Zhang Y P, Wei K P, Zhang Z, Hu W N, Zhang X M, Sun G G
School of Public Health, North China University of Science and Technology, Tangshan 063210, China.
Department of Oncologic Chemoradiotherapy, Tangshan Workers' Hospital, Tangshan 063001, China.
Zhonghua Zhong Liu Za Zhi. 2022 Apr 23;44(4):326-333. doi: 10.3760/cma.j.cn112152-20211214-00933.
To study the effects of dihydromyricetin (DMY) on the proliferation, apoptosis and epithelial mesenchymal transition (EMT) of esophageal squamous cell carcinoma (ESCC) cell KYSE150 and KYSE410. KYSE150 and KYSE410 cells were treated with different concentrations of DMY (0, 25, 50, 100, 150, 200 μmol/L) for 24 hours. The median inhibition concentration (IC) values of KYSE150 and KYSE410 were detected by cell counting kit-8 (CCK-8) method. Then 0.5‰ dimethyl sulfoxide (DMSO) was used as control group, dihydromyricetin (DMY), dihydromyricetin and transforming growth factor-β1 (DMY+ TGF-β1), transforming growth factor-β1 (TGF-β1) were used as experimental group. Cell proliferation and apoptosis rates were measured by clonal formation and flow cytometry. Transwell invasion and wound healing assay were used to detect cell invasion and migration. The protein expression levels of Caspase-3, Caspase-9, Bcl-2, Bax, Smad2/3, phosphorylation-Smad2/3 (p-Smad2/3) and Vimentin were detected by western blot. The IC values of DMY on KYSE410 and KYSE150 cells were 100.51 and 101.27 μmol/L. The clone formation numbers of KYSE150 and KYSE410 in DMY group [(0.53±0.03) and (0.31±0.03)] were lower than those in DMSO group [(1.00±0.10) and (1.00±0.05), <0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in DMY group [(1.84±0.22)% and (2.80±0.07)%] were higher than those in DMSO group [(1.00±0.18)% and (1.00±0.07)%, <0.05]. The invasion numbers of KYSE150 and KYSE410 cells in DMY group [(0.42±0.03) and (0.29±0.05)] were lower than those in DMSO group [(1.00±0.08) and (1.00±0.05), <0.05]. The migration rates of KYSE150 and KYSE410 cells in DMY group [(0.65±0.14)% and (0.40±0.17)%] were lower than those in DMSO group [(1.00±0.10)% and (1.00±0.08)%, <0.05]. The clone formation numbers of KYSE150 and KYSE410 in TGF-β1 group [(1.01±0.08) and (0.99±0.25)] were higher than those in DMY+ TGF-β1 group [(0.73±0.10) and (0.58±0.05), <0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in TGF-β1 group [(0.81±0.14)% and (1.18±0.10)%] were lower than those in DMY+ TGF-β1 group [(1.38±0.22)% and (1.85±0.04)%, <0.05]. The invasion numbers of KYSE150 and KYSE410 cells in TGF-β1 group [(1.19±0.11) and (1.39±0.11)] were higher than those in DMY+ TGF-β1 group [(0.93±0.09) and (0.93±0.05), <0.05]. The migration rates of KYSE150 and KYSE410 cells in TGF-β1 group [(1.87±0.19)% and (1.32±0.04)%] were higher than those in DMY+ TGF-β1 group [(0.86±0.16)% and (0.77±0.12)%, <0.05]. The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY group were higher than those in DMSO group, while the protein expression level of Bcl-2 was lower than that in DMSO group (<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in DMY group were lower than those in DMSO group (<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in TGF-β1 group were lower than those in DMY+ TGF-β1 group, and the protein expression level of Bcl-2 was higher than that in DMY+ TGF-β1 group (<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY+ TGF-β1 group were lower than those in DMY group, and the protein expression level of Bcl-2 was higher than that in DMY group (<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in TGF-β1 group were higher than those in DMY+ TGF-β1 group (<0.05). DMY can inhibit the proliferation and EMT of ESCC mediated by TGF-β1 and promote cell apoptosis.
研究二氢杨梅素（DMY）对食管鳞状细胞癌（ESCC）细胞KYSE150和KYSE410增殖、凋亡及上皮间质转化（EMT）的影响。将KYSE150和KYSE410细胞用不同浓度的DMY（0、25、50、100、150、200μmol/L）处理24小时。采用细胞计数试剂盒-8（CCK-8）法检测KYSE150和KYSE410的半数抑制浓度（IC）值。然后以0.5‰二甲基亚砜（DMSO）作为对照组，二氢杨梅素（DMY）、二氢杨梅素与转化生长因子-β1（DMY+TGF-β1）、转化生长因子-β1（TGF-β1）作为实验组。通过克隆形成和流式细胞术检测细胞增殖和凋亡率。采用Transwell侵袭实验和伤口愈合实验检测细胞侵袭和迁移能力。通过蛋白质印迹法检测Caspase-3、Caspase-9、Bcl-2、Bax、Smad2/3、磷酸化-Smad2/3（p-Smad2/3）和波形蛋白的蛋白表达水平。DMY对KYSE410和KYSE150细胞的IC值分别为100.51和101.27μmol/L。DMY组中KYSE150和KYSE410的克隆形成数[（0.53±0.03）和（0.31±0.03）]低于DMSO组[（1.00±0.10）和（1.00±0.05），<0.05]。DMY组中KYSE150和KYSE410细胞的凋亡率[（1.84±0.22）%和（2.80±0.07）%]高于DMSO组[（1.00±0.18）%和（1.00±0.07）%，<0.05]。DMY组中KYSE150和KYSE410细胞的侵袭数[（0.42±0.03）和（0.29±0.05）]低于DMSO组[（1.00±0.08）和（1.00±0.05），<0.05]。DMY组中KYSE150和KYSE410细胞的迁移率[（0.65±0.14）%和（0.40±0.17）%]低于DMSO组[（1.00±0.10）%和（1.00±0.08）%，<0.05]。TGF-β1组中KYSE150和KYSE410的克隆形成数[（1.01±0.08）和（0.99±0.25）]高于DMY+TGF-β1组[（0.73±0.10）和（0.58±0.05），<0.05]。TGF-β1组中KYSE150和KYSE410细胞的凋亡率[（0.81±0.14）%和（1.18±0.10）%]低于DMY+TGF-β1组[（1.38±0.22）%和（1.85±0.04）%，<0.05]。TGF-β1组中KYSE150和KYSE410细胞的侵袭数[（1.19±0.11）和（1.39±0.11）]高于DMY+TGF-β1组[（0.93±0.09）和（0.93±0.05），<0.05]。TGF-β1组中KYSE150和KYSE410细胞的迁移率[（1.87±0.19）%和（1.32±0.04）%]高于DMY+TGF-β1组[（0.86±0.16）%和（0.77±0.12）%，<0.05]。DMY组中KYSE150和KYSE410细胞的Bax、Caspase-3和Caspase-9蛋白表达水平高于DMSO组，而Bcl-2蛋白表达水平低于DMSO组（<0.05）。DMY组中KYSE150和KYSE410细胞的p-Smad2/3、Smad2/3和波形蛋白的蛋白表达水平低于DMSO组（<0.05）。TGF-β1组中KYSE150和KYSE410细胞的Bax、Caspase-3和Caspase-9蛋白表达水平低于DMY+TGF-β1组，而Bcl-2蛋白表达水平高于DMY+TGF-β1组（<0.05）。DMY+TGF-β1组中KYSE150和KYSE410细胞的Bax、Caspase-3和Caspase-9蛋白表达水平低于DMY组，而Bcl-2蛋白表达水平高于DMY组（<0.05）。TGF-β1组中KYSE150和KYSE410细胞的p-Smad2/3、Smad2/3和波形蛋白的蛋白表达水平高于DMY+TGF-β1组（<0.05）。二氢杨梅素可抑制TGF-β1介导的食管鳞状细胞癌增殖和EMT并促进细胞凋亡。
