长链酰基辅酶A合成酶3在肾透明细胞癌中抑制细胞增殖、迁移和侵袭的作用及其机制

Mechanism of long-chain Acyl-CoA synthetase 3 in inhibiting cell proliferation, migration and invasion in clear cell renal cell carcinoma

目的:探讨长链酰基辅酶A合成酶3(ACSL3)对肾透明细胞癌(ccRCC)细胞增殖、迁移、侵袭及脂质合成的影响。方法:使用肿瘤基因组图谱数据库分析ccRCC中ACSL3基因的表达情况。选取山西医科大学第一医院2021年10月至2022年10月手术切除的60对ccRCC和癌旁组织作为研究对象,利用荧光定量聚合酶链反应和免疫组织化学方法检测ccRCC组织中ACSL3表达水平;采用转染获得过表达ACSL3基因的细胞,采用细胞计数试剂盒(CCK-8)、平板克隆形成实验验证细胞的增殖能力;采用划痕实验和Transwell实验验证细胞的迁移、侵袭能力;采用流式细胞术检测细胞的凋亡能力;采用油红O实验检测细胞的脂质合成能力。两组间计量资料比较采用t检验,多组间比较采用方差分析。结果:ACSL3在ccRCC中的表达水平显著低于正常肾组织(0.42±0.20比1.01±0.16,t=17.946,P<0.05)。CCK-8结果显示转染96 h后,h-ACSL3组细胞吸光度(A)值显著低于对照组(1.095±0.047比1.295±0.025,t=8.393,P<0.05)。h-ACSL3组细胞形成的细胞集落数显著小于对照组[(309.00±93.30)个比(499.00±53.66)个,t=3.947,P<0.05]。h-ACSL3组划痕愈合率显著低于对照组[48 h:(45±7)%比(66±2)%,t=5.253,P<0.05]。h-ACSL3组穿过基底膜细胞数少于对照组[(1445.33±107.75)个比(1963.00±294.28)个,t=2.861,P<0.05]。h-ACSL3组细胞凋亡率高于对照组[(7.28±1.95)%比(3.53±0.24)%,t=-3.305,P<0.05]。h-ACSL3组细胞脂滴积累减少。结论:ACSL3在ccRCC中表达降低,ACSL3过表达会抑制ccRCC的增殖、迁移、侵袭能力,促进细胞凋亡,影响癌细胞中脂滴聚集。

Objective To investigate the effects of long-chain acyl-CoA synthetase 3(ACSL3)on the proliferation,migration,invasion and lipid synthesis of clear cell renal cell carcinoma(ccRCC)cells.Methods The TCGA database was used to compare the differential expression of ACSL3 in ccRCC.ccRCC and paraneoplastic tissues from the First Hospital of Shanxi Medical University were selected for the study,and the expression levels of ACSL3 in ccRCC tissues were detected using fluorescence quantitative polymerase chain reaction and immunohistochemistry.The cell counting kit(CCK-8)and plate clone formation assays were used to verify the proliferation ability of cells.Scratch assay and Transwell assay were used to verify the migration and invasion ability of cells.Flow cytometry was used to detect the apoptosis ability of cells.The oil red O assay was used to detect the lipid synthesis ability of cells.The t-test was used to compare the measurement data between two groups,and ANOVA was used to compare between multiple groups.Results ACSL3 was downregulated in ccRCC as compared with normal tissue[(0.42±0.20)vs.(1.01±0.16),t=17.946,P<0.05].CCK-8 results showed that after 96 h of transfection,the absorbance(A)values were lower in h-ACSL3 group than in control group[(1.095±0.047)vs.(1.295±0.025),t=8.393,P<0.05].The number of cell colonies in h-ACSL3 group was significantly less than that in control group[(309.00±93.30)vs.(499.00±53.66),t=3.947,P<0.05].The scratch healing rate in h-ACSL3 group was significantly lower than that in control group[48 h:(45±7)%vs.(66±2)%,t=5.253,P<0.05].The number of cells crossing the matrigel in h-ACSL3 group was less than in control group[(1445.33±107.75)vs.(1963.00±294.28),t=2.861,P<0.05].The apoptosis rate was significantly higher in h-ACSL3 group than in control group[(7.28±1.95)%vs.(3.53±0.24)%,t=-3.305,P<0.05].Overexpression of ACSL3 reduced the accumulation of abnormal lipid droplets in cells.Conclusion ACSL3 was downregulated in ccRCC.ACSL3 overexpression could inhibit the proliferation,migration and invasive ability,promote apoptosis,and affecte lipid droplet aggregation in ccRCC.