摘要
目的探讨辛二酰-双羟肟酸(suberoyl bis-hydroxamic acid,SBHA)对B16黑色素瘤增殖的影响及其抗肿瘤作用机制。方法将小鼠黑色素瘤B16细胞接种于培养瓶中进行培养,取对数期细胞进行试验,细胞增殖能力测试采用MTT法,Western Blot检测B16黑色素瘤细胞中Akt、p-Akt、PTEN 3种蛋白的表达。结果SBHA作用48h后,细胞活力随着SBHA浓度增加而下降,与空白对照组比较,差异有统计学意义(P<0.05)。SBHA作用72h后,对细胞活力的抑制作用明显比48h强,差异有统计学意义(P<0.05);给药72h药物浓度为200μmol/L时抑制作用达到最大化,IC50值为(43.50±0.98)μmol/L。结果表明,SBHA对黑色素瘤B16细胞增殖的抑制呈浓度和时间依赖关系(P<0.05);不同浓度的SBHA均对B16细胞Akt的表达具有抑制作用,与对照组比较,B16细胞Akt的表达量随着SBHA浓度的增加而呈现下降趋势(P<0.05),72h后B16细胞Akt的表达量下降趋势较48h后更明显,差异有统计学意义(P<0.05)。与对照组比较,B16细胞PTEN的表达量随着SBHA浓度的增加而呈现增长趋势,且差异有统计学意义(P<0.05);72h后B16细胞PTEN表达量的增长趋势较48h后更明显,差异有统计学意义(P<0.05)。不同浓度的SBHA均对B16细胞p-Akt的表达具有抑制作用,B16细胞p-Akt的表达量随着SBHA浓度的增加而呈现下降趋势,且差异有统计学意义(P<0.05)。结论 SBHA能够有效抑制细胞增殖,其机制可能是通过上调PTEN蛋白的表达并抑制Akt、p-Akt蛋白的表达实现。
Objective To investigate the effect of suberoyl bis-hydroxamic acid(SBHA)on the proliferation of B16melanoma and its anti-tumor mechanism.Methods The murine B16melanoma cells were inoculated into the culture bottle,and the logarithmic phase cells were tested.The cell proliferation was tested by MTT,and the expression of the three proteins Akt,p-Akt and PTEN in the B16melanoma cells were detected by Western Blot.Results The cell viability decreased with the increase of SBHA concentration after48-hour treatment with SBHA,and the difference was statistically significant when compared with the blank control group(P<0.05).The inhibition effect of the cell viability was significantly greater after72-hour treatment with SBHA than after48-hour treatment(P<0.05),and the inhibition effect was maximized when the drug concentration of SBHA was200μmol/L for72-hour treatment with the IC50value of(43.5±0.98)μmol/L.The inhibition effect of SBHA on the proliferation of B16melanoma cells was concentration-and time-dependent(P<0.05).Different concentrations of SBHA inhibited the expression of Akt in B16cells.Compared with the control group,the expression of Akt in B16cells decreased with the increase of SBHA concentration and the difference was statistically significant(P<0.05).The decrease of the Akt expression in B16cells was more significant after72hours than after48hours(P<0.05).The expression of PTEN in B16cells increased with the increase of SBHA concentration when compared with the control group,and the difference was statistically significant(P<0.05).The increase of the PTEN expression in B16cells was significantly greater after72hours than after48hours(P<0.05).Different concentrations of SBHA inhibited the expression of p-Akt in B16cells.The expression of p-Akt in B16cells decreased significantly with the increase of SBHA concentration(P<0.05).Conclusion SBHA can inhibit the cell proliferation effectively,and its mechanism may be performed by upregulating the expression of PTEN and inhibiting the expression of Akt and p-Akt.
作者
丁小洁
刘艳
陈燕
王理
熊芬
Ding Xiaojie;Liu Yan;Chen Yan;Wang Li;Xiong Fen(Department of Dermatology,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)
出处
《成都医学院学报》
CAS
2018年第6期658-661,共4页
Journal of Chengdu Medical College
基金
四川省医学科研项目资助(No:S17027)