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目的:观察组蛋白去乙酰化酶抑制剂辛二酰苯胺异羟肟酸(suberoylanilide hydroxamic acid,SAHA)体外对人卵巢癌SKOV3细胞转移潜能的影响,并探讨其可能机制。方法:体外应用SAHA作用于人卵巢癌SKOV3细胞,实验分为对照组、4μmol/L SAHA组、8μmol/L SAHA组和12μmol/L SAHA组,划痕实验检测SKOV3细胞迁移能力,Transwell侵袭实验检测SKOV3细胞侵袭能力。蛋白质印迹法检测SKOV3细胞内组蛋白H4乙酰化水平,实时定量PCR方法检测SKOV3细胞uPA和uPAR基因mRNA表达。结果:经SAHA作用48h后,对照组以及4、8和12μmol/L组细胞Ac-H4表达量分别为0.10±0.04、0.21±0.05、0.40±0.05和0.72±0.04,Ac-H4表达量随SAHA剂量浓度增加而增加,P<0.001;各组细胞划痕愈合率分别为(68.27±2.98)%、(35.19±2.97)%、(18.82±2.96)%和(10.24±2.98)%,划痕愈合率随SAHA剂量浓度增加而降低,P<0.001;各组细胞穿过Transwell滤膜的细胞数量分别为58.89±4.35、37.66±4.27、21.15±4.32和10.75±4.30,穿过Transwell滤膜的细胞数量随SAHA剂量浓度增加而减少,P<0.001;各组细胞uPAmRNA表达分别为22.48±1.05、15.40±1.02、8.62±1.05和5.27±1.08,随着SAHA剂量浓度增加而降低,P<0.001;各组细胞uPAR基因mRNA表达量分别为18.22±1.12、12.37±1.14、7.64±1.10和3.55±1.12,随SAHA剂量浓度增加而降低,P<0.001。结论:SAHA可抑制人卵巢癌SKOV3细胞迁移和侵袭能力,提高组蛋白乙酰化水平,降低uPA和uPAR基因表达可能是其作用机制。
Objective: To investigate the effect of suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, on the metastatic potential of human ovarian cancer SKOV3 cells in vitro and to explore its possible mechanism. Methods: SAHA was applied to human ovarian cancer cell line SKOV3 in vitro. The experiment was divided into control group, 4μmol / L SAHA group, 8μmol / L SAHA group and 12μmol / L SAHA group. Scratch assay was used to detect SKOV3 cell migration and Transwell invasion assay SKOV3 cell invasiveness. The histone H4 acetylation level in SKOV3 cells was detected by Western blotting, and the mRNA expression of uPA and uPAR in SKOV3 cells was detected by real-time quantitative PCR. Results: After SAHA for 48h, the expression of Ac-H4 in control group, 4, 8 and 12μmol / L groups were 0.10 ± 0.04, 0.21 ± 0.05, 0.40 ± 0.05 and 0.72 ± 0.04, respectively. (P <0.001). The healing rates of scratches in the groups were (68.27 ± 2.98)%, (35.19 ± 2.97)%, (18.82 ± 2.96)% and (10.24 ± 2.98)%, respectively P <0.001. The number of cells passing through the Transwell filter was 58.89 ± 4.35, 37.66 ± 4.27, 21.15 ± 4.32 and 10.75 ± 4.30, respectively. The number of cells passing through the Transwell filter (P <0.001). The expressions of uPA mRNA in each group were 22.48 ± 1.05,15.40 ± 1.02,8.62 ± 1.05 and 5.27 ± 1.08, respectively, and decreased with the increase of SAHA dose concentration (P <0.001) The expression of uPAR mRNA was 18.22 ± 1.12, 12.37 ± 1.14, 7.64 ± 1.10 and 3.55 ± 1.12, respectively, which decreased with the increase of SAHA dosage, P <0.001. Conclusion: SAHA can inhibit the migration and invasion of human ovarian cancer SKOV3 cells, increase the level of histone acetylation and decrease the expression of uPA and uPAR, which may be its mechanism of action.