中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2012年
11期
821-825
,共5页
韩建军%于金明%吴会勇%刘吉兵%宋宝%薛德文
韓建軍%于金明%吳會勇%劉吉兵%宋寶%薛德文
한건군%우금명%오회용%류길병%송보%설덕문
癌,肝细胞%复方斑蝥胶囊%细胞凋亡%免疫%小鼠
癌,肝細胞%複方斑蝥膠囊%細胞凋亡%免疫%小鼠
암,간세포%복방반모효낭%세포조망%면역%소서
Carcinoma,hepatocellular%Compound cantharides capsule%Apoptosis%Immunity%Mice
目的 探讨复方斑蝥胶囊对人HepG2215肝细胞癌移植瘤的抑制作用及其作用机制.方法 建立人HepG2215肝癌荷瘤小鼠模型,随机分为5组,A组为生理盐水对照组；B组为复方斑蝥胶囊组,其中B1组为12.5 mg·kg-1·d-1复方斑蝥胶囊,B2组为25 mg·kg-1 ·d-1复方斑蝥胶囊,B3组为37.5 mg·kg-1·d-1复方斑蝥胶囊；C组为环磷酰胺组(25 mg·kg-1·d-1).检测各组小鼠肿瘤体积、重量和外周血甲胎蛋白(AFP)浓度、HBV DNA水平；采用荧光定量逆转录聚合酶链反应(RT-PCR)检测肝癌凋亡相关基因mRNA的表达；采用TUNEL方法检测肿瘤细胞凋亡水平；采用流式细胞仪检测CD3+、CD19+、CD4+、CD8+的水平,免疫组化SP法检测肿瘤微血管密度(MVD).结果 用药结束后,B1组、B2组和B3组的抑瘤率分别为29.8％、38.7％和48.1％,C组抑瘤率为52.4％,各组间抑瘤率比较,差异有统计学意义(P＜0.05).A组、B1组、B2组、B3组和C组荷瘤小鼠的中位生存时间分别为(30.0±3.2)d、(49.0±5.1)d、(50.0±5.2)d、(57.5 ±6.5)d和(49.0±4.7)d,B3组小鼠中位生存时间明显高于其他各组(P＜0.05).A组、B1组、B2组、B3组和C组的小鼠血清AFP浓度分别为(492.7 ±48.5) ng/ml、(281.2 ±25.6) ng/ml、(194.3±18.7)ng/ml、(170.1±15.8) ng/ml和(138.7±12.5)ng/ml,C组可显著抑制AFP的表达.B1组、B2组、B3组和C组的HBV DNA抑制率分别为(46.0±5.1)％、(65.5±6.9)％、(81.3±7.8)％和(19.5±2.1)％,复方斑蝥胶囊可明显抑制HBV DNA复制,并有剂量依赖性.A组、B1组、B2组、B3组和C组的细胞凋亡率分别为(0.27±0.03)％、(7.18±2.12)％、(9.17±2.42)％、(11.27 ±3.03)％和(5.44±2.45)％,B3组的细胞凋亡率明显高于A组、B1组、B2组和C组(P＜0.05).B3组的bax mRNA表达水平明显高于C组(P＜0.05).复方斑蝥胶囊可明显下调bcl-2 mRNA表达,随着剂量的增加,对bcl-2 mRNA表达下调越明显,B3组的bcl-2 mRNA显著低于C组(P＜0.05).B3组的CD4+、CD8+、CD3+和CD19+水平明显高于A组和C组,差异有统计学意义(P＜0.05).B3组肝癌MVD明显小于A组和C组,差异有统计学意义(P＜0.05).结论 复方斑蝥胶囊可抑制HepG2215细胞中的HBV DNA复制,诱导肝癌细胞的凋亡；并通过提高免疫功能抑制肝癌细胞的生长,明显延长小鼠的中位生存时间.
目的 探討複方斑蝥膠囊對人HepG2215肝細胞癌移植瘤的抑製作用及其作用機製.方法 建立人HepG2215肝癌荷瘤小鼠模型,隨機分為5組,A組為生理鹽水對照組；B組為複方斑蝥膠囊組,其中B1組為12.5 mg·kg-1·d-1複方斑蝥膠囊,B2組為25 mg·kg-1 ·d-1複方斑蝥膠囊,B3組為37.5 mg·kg-1·d-1複方斑蝥膠囊；C組為環燐酰胺組(25 mg·kg-1·d-1).檢測各組小鼠腫瘤體積、重量和外週血甲胎蛋白(AFP)濃度、HBV DNA水平；採用熒光定量逆轉錄聚閤酶鏈反應(RT-PCR)檢測肝癌凋亡相關基因mRNA的錶達；採用TUNEL方法檢測腫瘤細胞凋亡水平；採用流式細胞儀檢測CD3+、CD19+、CD4+、CD8+的水平,免疫組化SP法檢測腫瘤微血管密度(MVD).結果 用藥結束後,B1組、B2組和B3組的抑瘤率分彆為29.8％、38.7％和48.1％,C組抑瘤率為52.4％,各組間抑瘤率比較,差異有統計學意義(P＜0.05).A組、B1組、B2組、B3組和C組荷瘤小鼠的中位生存時間分彆為(30.0±3.2)d、(49.0±5.1)d、(50.0±5.2)d、(57.5 ±6.5)d和(49.0±4.7)d,B3組小鼠中位生存時間明顯高于其他各組(P＜0.05).A組、B1組、B2組、B3組和C組的小鼠血清AFP濃度分彆為(492.7 ±48.5) ng/ml、(281.2 ±25.6) ng/ml、(194.3±18.7)ng/ml、(170.1±15.8) ng/ml和(138.7±12.5)ng/ml,C組可顯著抑製AFP的錶達.B1組、B2組、B3組和C組的HBV DNA抑製率分彆為(46.0±5.1)％、(65.5±6.9)％、(81.3±7.8)％和(19.5±2.1)％,複方斑蝥膠囊可明顯抑製HBV DNA複製,併有劑量依賴性.A組、B1組、B2組、B3組和C組的細胞凋亡率分彆為(0.27±0.03)％、(7.18±2.12)％、(9.17±2.42)％、(11.27 ±3.03)％和(5.44±2.45)％,B3組的細胞凋亡率明顯高于A組、B1組、B2組和C組(P＜0.05).B3組的bax mRNA錶達水平明顯高于C組(P＜0.05).複方斑蝥膠囊可明顯下調bcl-2 mRNA錶達,隨著劑量的增加,對bcl-2 mRNA錶達下調越明顯,B3組的bcl-2 mRNA顯著低于C組(P＜0.05).B3組的CD4+、CD8+、CD3+和CD19+水平明顯高于A組和C組,差異有統計學意義(P＜0.05).B3組肝癌MVD明顯小于A組和C組,差異有統計學意義(P＜0.05).結論 複方斑蝥膠囊可抑製HepG2215細胞中的HBV DNA複製,誘導肝癌細胞的凋亡；併通過提高免疫功能抑製肝癌細胞的生長,明顯延長小鼠的中位生存時間.
목적 탐토복방반모효낭대인HepG2215간세포암이식류적억제작용급기작용궤제.방법 건립인HepG2215간암하류소서모형,수궤분위5조,A조위생리염수대조조；B조위복방반모효낭조,기중B1조위12.5 mg·kg-1·d-1복방반모효낭,B2조위25 mg·kg-1 ·d-1복방반모효낭,B3조위37.5 mg·kg-1·d-1복방반모효낭；C조위배린선알조(25 mg·kg-1·d-1).검측각조소서종류체적、중량화외주혈갑태단백(AFP)농도、HBV DNA수평；채용형광정량역전록취합매련반응(RT-PCR)검측간암조망상관기인mRNA적표체；채용TUNEL방법검측종류세포조망수평；채용류식세포의검측CD3+、CD19+、CD4+、CD8+적수평,면역조화SP법검측종류미혈관밀도(MVD).결과 용약결속후,B1조、B2조화B3조적억류솔분별위29.8％、38.7％화48.1％,C조억류솔위52.4％,각조간억류솔비교,차이유통계학의의(P＜0.05).A조、B1조、B2조、B3조화C조하류소서적중위생존시간분별위(30.0±3.2)d、(49.0±5.1)d、(50.0±5.2)d、(57.5 ±6.5)d화(49.0±4.7)d,B3조소서중위생존시간명현고우기타각조(P＜0.05).A조、B1조、B2조、B3조화C조적소서혈청AFP농도분별위(492.7 ±48.5) ng/ml、(281.2 ±25.6) ng/ml、(194.3±18.7)ng/ml、(170.1±15.8) ng/ml화(138.7±12.5)ng/ml,C조가현저억제AFP적표체.B1조、B2조、B3조화C조적HBV DNA억제솔분별위(46.0±5.1)％、(65.5±6.9)％、(81.3±7.8)％화(19.5±2.1)％,복방반모효낭가명현억제HBV DNA복제,병유제량의뢰성.A조、B1조、B2조、B3조화C조적세포조망솔분별위(0.27±0.03)％、(7.18±2.12)％、(9.17±2.42)％、(11.27 ±3.03)％화(5.44±2.45)％,B3조적세포조망솔명현고우A조、B1조、B2조화C조(P＜0.05).B3조적bax mRNA표체수평명현고우C조(P＜0.05).복방반모효낭가명현하조bcl-2 mRNA표체,수착제량적증가,대bcl-2 mRNA표체하조월명현,B3조적bcl-2 mRNA현저저우C조(P＜0.05).B3조적CD4+、CD8+、CD3+화CD19+수평명현고우A조화C조,차이유통계학의의(P＜0.05).B3조간암MVD명현소우A조화C조,차이유통계학의의(P＜0.05).결론 복방반모효낭가억제HepG2215세포중적HBV DNA복제,유도간암세포적조망；병통과제고면역공능억제간암세포적생장,명현연장소서적중위생존시간.
Objective To investigate the inhibitory effect of compound cantharides capsules on the proliferation of xenografts of human hepatocellular carcinoma HepG2215 in mice and their mechanism of action.Methods One hundred healthy Balb/c mice (5-week old,male: female 1:1) were used in this study.Mouse models of human HepG2215 hepatocarcinoma were established.The tumor-bearing mice were divided into five groups randomly.The control group A received daily intragastric administration of physiologic saline.The intervention groups B1,B2 and B3 were treated with compound cantharides capsule in a dose of12.5 mg· kg-1 · d-1,25 mg· kg-1 · d-1 and 37.5 mg · kg-1 · d-1,respectively,for 10 consecutive days.The group C had intraperitoneal injection of cyclophosphamide (25 mg · kg-1 · d-1) for l0 consecutive days.The mice were sacrificed after the completion of administration.The tumors were taken out,the tumor volume was measured,the inhibitory rate of body weight was calculated,and the serum AFP concentration and the level of HBV DNA were determined.The survival of each group mice was analyzed.The levels of mRNA expression of apoptosis-related genes were assayed by quantitative RT-PCR.Apoptosis in the tumor cells was assayed with TUNEL staining.Flow cytometry was used to detect the levels of CD3 +,CD19+,CD4+ and CD8+,and microvessel density (MVD) of the tumors was assessed by immunohistochemistry.Results After completion of the treatment,the inhibition rate of tumor growth of the groups B1,B2 and B3 was 29.8％,38.7％ and 48.1％,respectively,and that of the group C was 52.4％,with a significant difference among the groups (P ＜0.05).The median survival time of the groups A,B1,B2,B3 and C was (30.0±3.2) days,(49.0 ±5.1) days,(50.0 ±5.2) days,(57.5 ±6.5) days and (49.0±4.7) days,respectively.The median survival time of the group B3 was significantly longer than that of other groups (P＜0.05).The serum AFP level in the groups A,B1,B2,B3 and C was (492.7 ± 48.5) ng/ml,(281.2 ±25.6) ng/ml,(194.3 ±18.7) ng/ml,(170.1 ±15.8) ng/ml and (138.7 ±12.5) ng/ml,respectively,indicating that it was significantly inhibited in the group C.The inhibition rate of HBV DNA replication of the groups B1,B2,B3 and C was (46.0 ±5.1)％,(65.5 ±6.9)％,(81.3 ±7.8)％ and (19.5 ± 2.1)％,respectively,showing that compound cantharides capsules inhibited HBV DNA replication in a dose-dependent manner.The apoptosis rate of the groups A,B1,B2,B3 and C was (0.27 ± 0.03)％,(7.18±2.12)％,(9.17 ±2.42)％,(11.27±3.03)％ and (5.44 ±2.45)％,respectively,and that of the group B3 was significantly higher than that of the groups A,B1,B2 and C (P ＜ 0.05).The expression level of bax mRNA was significantly higher than that of the group C (P ＜ 0.05).The drug could significantly decrease the bcl-2 mRNA expression level,more remarkably along with the increasing dose of cantharides,and it was significantly lower than that in the group C (P ＜ 0.05).The levels of CD4 +,CD8 +,CD3 + and CD19 + were significantly higher than that in the groups A and C (P ＜0.05).The value of MVD of the group B3 was significantly lower that that of groups A and C (P ＜0.05).Conclusion Compound cantharides capsules may inhibit the replication of HBV DNA in HepG2215 cells,inducing apoptosis in the tumor cells,enhancing the immune function to inhibit the growth of liver cancer cells in mice,and significantly prolong the median survival time of tumor-bearing mice.
