中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
2期
322-326
,共5页
刘洋%刘振操%邱少敏%刘琳
劉洋%劉振操%邱少敏%劉琳
류양%류진조%구소민%류림
结肠癌%组蛋白去乙酰化酶抑制剂%脱噬作用%裸鼠
結腸癌%組蛋白去乙酰化酶抑製劑%脫噬作用%裸鼠
결장암%조단백거을선화매억제제%탈서작용%라서
Colorectal cancer%Histone deacetylase inhibitor%Apoptosis%Nude mice
目的 观察组蛋白去乙酰化酶抑制剂(HDACi)西达本胺(CS055)对人结肠癌裸鼠皮下移植瘤的抑制作用,并探讨其机制.方法 成功建立人结肠癌裸鼠皮下移植瘤模型,将55只裸鼠随机分成5组,每组11只,分别予以CS055 10.0、5.0、2.5 mg/(kg·d)灌胃和5-氟尿嘧啶(5-Fu)25.0 mg/(kg·d)腹腔注射,以及阴性对照药物0.1 ml/10 g灌胃干预,连续给药21 d.治疗结束后,测量各组瘤体积及瘤重,计算瘤体积抑制率及瘤重抑制率.取瘤组织行苏木素-伊红(HE)染色,通过透射电镜观察肿瘤细胞凋亡,Westem blot检测蛋白去乙酰化酶l(HDACl)、HDAC2、组蛋白-H3(ac-H3)、p21、周期蛋白依赖性激酶4(CDK4)、半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3的表达.结果 与阴性对照药物比较,高剂量组、中剂量组、低剂量组、5-Fu组抑瘤率分别为15.36％(P＞0.05)、40.27％ (P＜ 0.01)、12.29％ (P＞0.05)、55.63％ (P＜ 0.01);瘤体积抑制率分别为15.48％ (P＞ 0.05)、40.47％ (P＜0.01)、12.37％ (P＞0.05)、55.74％(P＜0.01).CS055能抑制HDAC1和HDAC2的表达,提高ac-H3的乙酰化水平;同时诱导p21、Caspase-3的表达,下调CDK4的表达.结论 CS055能抑制人结肠癌裸鼠皮下移植瘤的生长,其机制可能为抑制HDAC1和HDAC2的表达,提高ac-H3的乙酰化水平,通过上调p21的蛋白表达,下调CDK4的蛋白表达,诱导结肠癌细胞周期阻滞;同时上调Caspase-3的蛋白表达,诱导结肠癌细胞凋亡.
目的 觀察組蛋白去乙酰化酶抑製劑(HDACi)西達本胺(CS055)對人結腸癌裸鼠皮下移植瘤的抑製作用,併探討其機製.方法 成功建立人結腸癌裸鼠皮下移植瘤模型,將55隻裸鼠隨機分成5組,每組11隻,分彆予以CS055 10.0、5.0、2.5 mg/(kg·d)灌胃和5-氟尿嘧啶(5-Fu)25.0 mg/(kg·d)腹腔註射,以及陰性對照藥物0.1 ml/10 g灌胃榦預,連續給藥21 d.治療結束後,測量各組瘤體積及瘤重,計算瘤體積抑製率及瘤重抑製率.取瘤組織行囌木素-伊紅(HE)染色,通過透射電鏡觀察腫瘤細胞凋亡,Westem blot檢測蛋白去乙酰化酶l(HDACl)、HDAC2、組蛋白-H3(ac-H3)、p21、週期蛋白依賴性激酶4(CDK4)、半胱氨酰天鼕氨痠特異性蛋白酶(Caspase)-3的錶達.結果 與陰性對照藥物比較,高劑量組、中劑量組、低劑量組、5-Fu組抑瘤率分彆為15.36％(P＞0.05)、40.27％ (P＜ 0.01)、12.29％ (P＞0.05)、55.63％ (P＜ 0.01);瘤體積抑製率分彆為15.48％ (P＞ 0.05)、40.47％ (P＜0.01)、12.37％ (P＞0.05)、55.74％(P＜0.01).CS055能抑製HDAC1和HDAC2的錶達,提高ac-H3的乙酰化水平;同時誘導p21、Caspase-3的錶達,下調CDK4的錶達.結論 CS055能抑製人結腸癌裸鼠皮下移植瘤的生長,其機製可能為抑製HDAC1和HDAC2的錶達,提高ac-H3的乙酰化水平,通過上調p21的蛋白錶達,下調CDK4的蛋白錶達,誘導結腸癌細胞週期阻滯;同時上調Caspase-3的蛋白錶達,誘導結腸癌細胞凋亡.
목적 관찰조단백거을선화매억제제(HDACi)서체본알(CS055)대인결장암라서피하이식류적억제작용,병탐토기궤제.방법 성공건립인결장암라서피하이식류모형,장55지라서수궤분성5조,매조11지,분별여이CS055 10.0、5.0、2.5 mg/(kg·d)관위화5-불뇨밀정(5-Fu)25.0 mg/(kg·d)복강주사,이급음성대조약물0.1 ml/10 g관위간예,련속급약21 d.치료결속후,측량각조류체적급류중,계산류체적억제솔급류중억제솔.취류조직행소목소-이홍(HE)염색,통과투사전경관찰종류세포조망,Westem blot검측단백거을선화매l(HDACl)、HDAC2、조단백-H3(ac-H3)、p21、주기단백의뢰성격매4(CDK4)、반광안선천동안산특이성단백매(Caspase)-3적표체.결과 여음성대조약물비교,고제량조、중제량조、저제량조、5-Fu조억류솔분별위15.36％(P＞0.05)、40.27％ (P＜ 0.01)、12.29％ (P＞0.05)、55.63％ (P＜ 0.01);류체적억제솔분별위15.48％ (P＞ 0.05)、40.47％ (P＜0.01)、12.37％ (P＞0.05)、55.74％(P＜0.01).CS055능억제HDAC1화HDAC2적표체,제고ac-H3적을선화수평;동시유도p21、Caspase-3적표체,하조CDK4적표체.결론 CS055능억제인결장암라서피하이식류적생장,기궤제가능위억제HDAC1화HDAC2적표체,제고ac-H3적을선화수평,통과상조p21적단백표체,하조CDK4적단백표체,유도결장암세포주기조체;동시상조Caspase-3적단백표체,유도결장암세포조망.
Objective To explore the inhibitory effect and its mechanism of CS055 [a selective histone deacetylase inhibitor (HDACi)] on human colon carcinoma in nude mice.Methods The xenograft model was established successfully.Fifty-five mice were divided into five groups and eleven blocks by randomized complete-block design.The mice were respectively garaged by CS055 10.0 mg/(kg·d) in the first group,5 mg/(kg·d) in the second group and 2.5 mg/(kg·d) in the third group,while the mice in fourth group were treated by intraperitoneal injection of 5-fluorouracil in a dose of 25.0 mg/(kg·d) and those in the fifth group were gavaged by negative-control-drug 0.1 ml/(10 g·d) as negative control group.After treatment,all nude mice were killed,and all the subcutaneous transplanted tumors were stripped and measured by calipers and scales.Inhibitory rate of tumor weight and tumor volume was calculated.Tissue hematoxylin and eosin (HE) staining and transmission electron microscopy detection were used to detect the tumor cell apoptosis.Western blotting was used to detect the expression of histone deacetylase 1 (HDAC1),HDAC2,acetylated histone-H3 (ac-H3),p21,cyclin dependent kinase 4 (CDK4),and cysteinyl aspartate-specific protease(Caspase)-3.Results Compared to negative control group,the respective inhibition rate of tumor weight in high-dose group,mediate-dose group,low-dose group,and5-fluorouracil group was 15.36％ (P＞0.05),40.27％ (P＜0.01),12.29％ (P＞ 0.05) and 55.63％ (P＜ 0.01) respectively,and the respective inhibitory rate of tumor volume was 15.48％ (P ＞ 0.05),40.47％ (P ＜ 0.01),12.37％ (P ＞ 0.05) and 55.74％ (P ＜ 0.01) respectively.CS055 could significantly inhibit the expression of HDAC1 and HDAC2,upregulate the level of acetylation of histonc H3 and the expression of p21,Caspase-3,while downregulate the expression of CDK4 protein.Conclusion CS055 can inhibit the growth of human colon carcinoma xenograft tumor probably by inhibiting the expression of HDAC1 and HDAC2,upregulating the level of histone H3 acetylation and the expression of p21 protein,downregulating the expression of CDK4 protein,which promotes tumor cell cycle arrest,and upregulating the expression of Caspase-3 protein,which induces LoVo cells apoptosis.
