中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
4期
256-261
,共6页
祝丽丽%胡万乐%张林君%俞志高%黄崇杰%姜明哲%滕明星%刘建录%刘长宝
祝麗麗%鬍萬樂%張林君%俞誌高%黃崇傑%薑明哲%滕明星%劉建錄%劉長寶
축려려%호만악%장림군%유지고%황숭걸%강명철%등명성%류건록%류장보
结直肠肿瘤%辣椒素%凋亡%热休克蛋白类%caspase-3%Cyt-C
結直腸腫瘤%辣椒素%凋亡%熱休剋蛋白類%caspase-3%Cyt-C
결직장종류%랄초소%조망%열휴극단백류%caspase-3%Cyt-C
Colorectal neoplasms%Capsaicin%Apoptosis%Heat-shock proteins%Caspase-3%Cyt-C
目的 探讨辣椒素对结肠癌HT-29细胞裸鼠皮下移植瘤的抑瘤作用及其机制.方法 建立结肠癌HT-29细胞裸鼠皮下移植瘤模型,采用原位末端脱氧核苷酸转移酶标记法(TUNEL法)检测移植瘤的细胞凋亡情况,采用免疫组织化学染色法和Western blot检测肿瘤组织中热休克蛋白27(HSP27)、细胞色素C(Cyt-C)和活化caspase-3蛋白的表达.结果 与生理盐水组(0.9％氯化钠注射液,NS组)比较,中剂量辣椒素组(10 mg·kg-1·d-1,C10组)和高剂量辣椒素组(20 mg·kg-1·d-1,C20组)移植瘤生长速度缓慢.TUNEL法检测结果显示,低剂量辣椒素组(5 mg· kg-1·d-1,C5组)和C10组积分吸光度值分别为2532.14±578.11和6364.03±1137.98,均高于NS组(760.12 ±238.05,均P ＜0.05),C20组积分吸光度值(15743.96±1855.95)高于C5组、C10组和NS组(均P＜0.01).免疫组化结果显示,NS组肿瘤组织中HSP27蛋白呈强阳性表达,其表达随着辣椒素剂量的增加而减弱,而活化的caspase-3和Cyt-C呈弱阳性表达,其表达随着辣椒素剂量的增加而增强.Western blot 检测结果显示,C5组和C10组HSP27蛋白的相对表达水平分别为0.73 ±0.05和0.41 ±0.03,均低于NS组(均P＜0.05),C20组HSP27蛋白的相对表达水平(0.22±0.06)均低于NS组、C5组和C10组(均P＜0.01).C5组活化的caspase-3和Cyt-C蛋白的相对表达水平分别为2.57 ±0.34和2.03±0.38,均高于NS组(均P＜0.05).C10组活化的caspase-3和Cyt-C蛋白的相对表达水平分别为4.23±0.45和3.13 ±0.44,均高于NS组(均P＜0.05),C20组活化的caspase-3和Cyt-C蛋白的相对表达水平分别为5.78 ±0.48和4.92 ±0.52,均高于NS组、C5组和C10组(均P＜0.01).结论 辣椒素可抑制结肠癌HT-29细胞裸鼠皮下移植瘤的生长,诱导细胞凋亡,其作用机制可能与抑制HT-29细胞中HSP27蛋白的表达、促进活化的caspase-3、Cyt-C蛋白的表达有关.
目的 探討辣椒素對結腸癌HT-29細胞裸鼠皮下移植瘤的抑瘤作用及其機製.方法 建立結腸癌HT-29細胞裸鼠皮下移植瘤模型,採用原位末耑脫氧覈苷痠轉移酶標記法(TUNEL法)檢測移植瘤的細胞凋亡情況,採用免疫組織化學染色法和Western blot檢測腫瘤組織中熱休剋蛋白27(HSP27)、細胞色素C(Cyt-C)和活化caspase-3蛋白的錶達.結果 與生理鹽水組(0.9％氯化鈉註射液,NS組)比較,中劑量辣椒素組(10 mg·kg-1·d-1,C10組)和高劑量辣椒素組(20 mg·kg-1·d-1,C20組)移植瘤生長速度緩慢.TUNEL法檢測結果顯示,低劑量辣椒素組(5 mg· kg-1·d-1,C5組)和C10組積分吸光度值分彆為2532.14±578.11和6364.03±1137.98,均高于NS組(760.12 ±238.05,均P ＜0.05),C20組積分吸光度值(15743.96±1855.95)高于C5組、C10組和NS組(均P＜0.01).免疫組化結果顯示,NS組腫瘤組織中HSP27蛋白呈彊暘性錶達,其錶達隨著辣椒素劑量的增加而減弱,而活化的caspase-3和Cyt-C呈弱暘性錶達,其錶達隨著辣椒素劑量的增加而增彊.Western blot 檢測結果顯示,C5組和C10組HSP27蛋白的相對錶達水平分彆為0.73 ±0.05和0.41 ±0.03,均低于NS組(均P＜0.05),C20組HSP27蛋白的相對錶達水平(0.22±0.06)均低于NS組、C5組和C10組(均P＜0.01).C5組活化的caspase-3和Cyt-C蛋白的相對錶達水平分彆為2.57 ±0.34和2.03±0.38,均高于NS組(均P＜0.05).C10組活化的caspase-3和Cyt-C蛋白的相對錶達水平分彆為4.23±0.45和3.13 ±0.44,均高于NS組(均P＜0.05),C20組活化的caspase-3和Cyt-C蛋白的相對錶達水平分彆為5.78 ±0.48和4.92 ±0.52,均高于NS組、C5組和C10組(均P＜0.01).結論 辣椒素可抑製結腸癌HT-29細胞裸鼠皮下移植瘤的生長,誘導細胞凋亡,其作用機製可能與抑製HT-29細胞中HSP27蛋白的錶達、促進活化的caspase-3、Cyt-C蛋白的錶達有關.
목적 탐토랄초소대결장암HT-29세포라서피하이식류적억류작용급기궤제.방법 건립결장암HT-29세포라서피하이식류모형,채용원위말단탈양핵감산전이매표기법(TUNEL법)검측이식류적세포조망정황,채용면역조직화학염색법화Western blot검측종류조직중열휴극단백27(HSP27)、세포색소C(Cyt-C)화활화caspase-3단백적표체.결과 여생리염수조(0.9％록화납주사액,NS조)비교,중제량랄초소조(10 mg·kg-1·d-1,C10조)화고제량랄초소조(20 mg·kg-1·d-1,C20조)이식류생장속도완만.TUNEL법검측결과현시,저제량랄초소조(5 mg· kg-1·d-1,C5조)화C10조적분흡광도치분별위2532.14±578.11화6364.03±1137.98,균고우NS조(760.12 ±238.05,균P ＜0.05),C20조적분흡광도치(15743.96±1855.95)고우C5조、C10조화NS조(균P＜0.01).면역조화결과현시,NS조종류조직중HSP27단백정강양성표체,기표체수착랄초소제량적증가이감약,이활화적caspase-3화Cyt-C정약양성표체,기표체수착랄초소제량적증가이증강.Western blot 검측결과현시,C5조화C10조HSP27단백적상대표체수평분별위0.73 ±0.05화0.41 ±0.03,균저우NS조(균P＜0.05),C20조HSP27단백적상대표체수평(0.22±0.06)균저우NS조、C5조화C10조(균P＜0.01).C5조활화적caspase-3화Cyt-C단백적상대표체수평분별위2.57 ±0.34화2.03±0.38,균고우NS조(균P＜0.05).C10조활화적caspase-3화Cyt-C단백적상대표체수평분별위4.23±0.45화3.13 ±0.44,균고우NS조(균P＜0.05),C20조활화적caspase-3화Cyt-C단백적상대표체수평분별위5.78 ±0.48화4.92 ±0.52,균고우NS조、C5조화C10조(균P＜0.01).결론 랄초소가억제결장암HT-29세포라서피하이식류적생장,유도세포조망,기작용궤제가능여억제HT-29세포중HSP27단백적표체、촉진활화적caspase-3、Cyt-C단백적표체유관.
Objective To evaluate the effect of capsaicin on nude mice xenografted with colorectal carcinoma cells,and to explore its mechanism of action.Methods A nude mouse model of colorectal cancer was established by subcutaneous inoculation of human colorectal carcinoma HT-29 cells.Terminal deoxynucleotidyl transferase-mediated nicked labeling assay (TUNEL) was undertaken to detect the cell proliferation and apoptosis in the xenograft tissue in nude mice.Immunohistochemical (IHC) staining and Western blot were used to detect the expression of HSP27,Cyt-C and active caspase-3.Results The tumor growth of the groups C10 and C20 was significantly slower than that of the group NS.The integrated optical density (IOD) of both the group C5 (2532.14 ± 578.11) and group C10 (6364.03 ± 1137.98) was significantly higher than that of the group NS (760.12 ± 238.05),(P ＜ 0.05).The integrated optical density (IOD) of the group C20 was (15743.96 ± 1855.95),significantly higher than that of the groups C10,C5 and NS (all were P ＜ 0.01).Immunohistochemistry showed that the cytoplasmic expression of HSP27 was strongly positive in the group NS,and significantly reduced with the increasing dose of capsaicin in the treated groups.The expression of active caspase-3 and Cyt-C in the group NS was weakly positive,and was significantly increased with the increasing dose of capsaicin in the groups C5 and C10 (P ＜0.05),and the expression of active caspase-3 and Cyt-C of the group C20 was significantly higher than that of the groups C5,C10 and NS (P ＜0.01).Western blot analysis showed that both the expressions of HSP27 of the group C5 (0.73 ±0.05) and the group C10 (0.41 ±0.03) were significantly lower than that of the group NS (P ＜ 0.05).The expression of HSP27 of the group C20 (0.22 ± 0.06) was significantly lower than that of the groups C5,C10 and NS (P ＜0.01).The expressions of active-caspase-3 and Cyt-C in the group C5 were (2.57 ± 0.34) and (2.03 ± 0.38),significantly higher than those of the group NS (P ＜0.05).The expressions of active-caspase-3 and Cyt-C in the group C10 were (4.23 ±0.45) and (3.13 ± 0.44),also significantly higher than those of the group NS (P ＜ 0.05).The expressions of active-caspase-3 and Cyt-C in the group C20 were (5.78 ± 0.48) and (4.92 ± 0.52),significantly higher than those of the group C5,C10 and NS (P ＜0.01).TUNEL analysis showed that there was a significant difference of cell apoptosis in comparison of each two groups.The higher dose of capsaicin was used,the more apoptosis was observed.Conclusions Capsaicin can significantly inhibit the tumor growth and induce cell apoptosis in the colorectal carcinoma xenograft in nude mice.Its mechanism of action is possibly related with the downregulation of HSP27 expression and up-regulation of expression of active caspase-3 and Cyt-C in the colorectal carcinoma xenograft in nude mice.
