中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
32期
5749-5756
,共8页
许峰%张雷%潘晋坤%薛利利%赵晓燕%李宝平
許峰%張雷%潘晉坤%薛利利%趙曉燕%李寶平
허봉%장뢰%반진곤%설리리%조효연%리보평
干细胞%骨髓干细胞%骨髓间充质干细胞%A549荷瘤小鼠肺癌%肿瘤相关成纤维细胞%间质纤维%免疫组化%α平滑肌收缩蛋白%管内皮生长因子%肝细胞生长因子%白细胞介素6%肌糖蛋白C%干细胞图片文章
榦細胞%骨髓榦細胞%骨髓間充質榦細胞%A549荷瘤小鼠肺癌%腫瘤相關成纖維細胞%間質纖維%免疫組化%α平滑肌收縮蛋白%管內皮生長因子%肝細胞生長因子%白細胞介素6%肌糖蛋白C%榦細胞圖片文章
간세포%골수간세포%골수간충질간세포%A549하류소서폐암%종류상관성섬유세포%간질섬유%면역조화%α평활기수축단백%관내피생장인자%간세포생장인자%백세포개소6%기당단백C%간세포도편문장
背景：肿瘤被认为是一种特殊的不愈合创伤，骨髓间充质干细胞通过向肿瘤组织归巢和向间质成分分化，参与肿瘤间质重构，从而改变肿瘤微环境，影响肿瘤的生长和转移。目的：在 A549肺癌荷瘤小鼠模型上证实骨髓间充质干细胞参与其损伤修复，探讨骨髓间充质干细胞参与肿瘤组织修复的机制。方法：体外分离、培养人骨髓间充质干细胞，使用流式细胞术鉴定，制造 A549肺癌荷瘤小鼠模型。实验组采用瘤周注射人骨髓间充质干细胞，对照组注射等量 PBS，对比观察动物生活情况，肿瘤生长大小。4周后取材，苏木精-伊红染色对比观察肿瘤组织，Masson染色对比胶原纤维含量，RT-PCR检测两组α平滑肌收缩蛋白的表达，免疫组织化学检测两组成纤维细胞特异蛋白、成纤维细胞活化蛋白的表达情况，反映两组肿瘤组织的间质纤维的程度。免疫组织化学方法对比两组中血管内皮生长因子、肝细胞生长因子、白细胞介素6、肌糖蛋白C的表达高低。结果与结论：骨髓间充质干细胞促进荷瘤小鼠肿瘤的生长，实验组肿瘤组织生长速度明显快于对照组(P <0.05)。RT-PCR检测α平滑肌收缩蛋白的表达：与对照组比较，实验组α平滑肌收缩蛋白 mRNA的表达水平显著升高。免疫组织化学方法检测实验组肿瘤组织中 TAFs 标志物：成纤维细胞特异蛋白、成纤维细胞活化蛋白的表达，IHC检测血管内皮生长因子、肝细胞生长因子、白细胞介素6、肌糖蛋白C的表达明显高于对照组，差异有显著性意义(P<0.05)。说明骨髓间充质干细胞在肿瘤微环境中向成纤维细胞方向分化，参与肿瘤间质的形成和构建，分泌血管内皮生长因子、肝细胞生长因子、白细胞介素6、肌糖蛋白C等促进肿瘤的生长修复。
揹景：腫瘤被認為是一種特殊的不愈閤創傷，骨髓間充質榦細胞通過嚮腫瘤組織歸巢和嚮間質成分分化，參與腫瘤間質重構，從而改變腫瘤微環境，影響腫瘤的生長和轉移。目的：在 A549肺癌荷瘤小鼠模型上證實骨髓間充質榦細胞參與其損傷脩複，探討骨髓間充質榦細胞參與腫瘤組織脩複的機製。方法：體外分離、培養人骨髓間充質榦細胞，使用流式細胞術鑒定，製造 A549肺癌荷瘤小鼠模型。實驗組採用瘤週註射人骨髓間充質榦細胞，對照組註射等量 PBS，對比觀察動物生活情況，腫瘤生長大小。4週後取材，囌木精-伊紅染色對比觀察腫瘤組織，Masson染色對比膠原纖維含量，RT-PCR檢測兩組α平滑肌收縮蛋白的錶達，免疫組織化學檢測兩組成纖維細胞特異蛋白、成纖維細胞活化蛋白的錶達情況，反映兩組腫瘤組織的間質纖維的程度。免疫組織化學方法對比兩組中血管內皮生長因子、肝細胞生長因子、白細胞介素6、肌糖蛋白C的錶達高低。結果與結論：骨髓間充質榦細胞促進荷瘤小鼠腫瘤的生長，實驗組腫瘤組織生長速度明顯快于對照組(P <0.05)。RT-PCR檢測α平滑肌收縮蛋白的錶達：與對照組比較，實驗組α平滑肌收縮蛋白 mRNA的錶達水平顯著升高。免疫組織化學方法檢測實驗組腫瘤組織中 TAFs 標誌物：成纖維細胞特異蛋白、成纖維細胞活化蛋白的錶達，IHC檢測血管內皮生長因子、肝細胞生長因子、白細胞介素6、肌糖蛋白C的錶達明顯高于對照組，差異有顯著性意義(P<0.05)。說明骨髓間充質榦細胞在腫瘤微環境中嚮成纖維細胞方嚮分化，參與腫瘤間質的形成和構建，分泌血管內皮生長因子、肝細胞生長因子、白細胞介素6、肌糖蛋白C等促進腫瘤的生長脩複。
배경：종류피인위시일충특수적불유합창상，골수간충질간세포통과향종류조직귀소화향간질성분분화，삼여종류간질중구，종이개변종류미배경，영향종류적생장화전이。목적：재 A549폐암하류소서모형상증실골수간충질간세포삼여기손상수복，탐토골수간충질간세포삼여종류조직수복적궤제。방법：체외분리、배양인골수간충질간세포，사용류식세포술감정，제조 A549폐암하류소서모형。실험조채용류주주사인골수간충질간세포，대조조주사등량 PBS，대비관찰동물생활정황，종류생장대소。4주후취재，소목정-이홍염색대비관찰종류조직，Masson염색대비효원섬유함량，RT-PCR검측량조α평활기수축단백적표체，면역조직화학검측량조성섬유세포특이단백、성섬유세포활화단백적표체정황，반영량조종류조직적간질섬유적정도。면역조직화학방법대비량조중혈관내피생장인자、간세포생장인자、백세포개소6、기당단백C적표체고저。결과여결론：골수간충질간세포촉진하류소서종류적생장，실험조종류조직생장속도명현쾌우대조조(P <0.05)。RT-PCR검측α평활기수축단백적표체：여대조조비교，실험조α평활기수축단백 mRNA적표체수평현저승고。면역조직화학방법검측실험조종류조직중 TAFs 표지물：성섬유세포특이단백、성섬유세포활화단백적표체，IHC검측혈관내피생장인자、간세포생장인자、백세포개소6、기당단백C적표체명현고우대조조，차이유현저성의의(P<0.05)。설명골수간충질간세포재종류미배경중향성섬유세포방향분화，삼여종류간질적형성화구건，분비혈관내피생장인자、간세포생장인자、백세포개소6、기당단백C등촉진종류적생장수복。
BACKGROUND:Tumor has been considered as a specific nonhealing trauma. Bone marrow mesenchymal stem cel s participate in tumor mesenchymal reconstitution by tumor tissue homing and differentiation into mesenchyme, resulting in changing tumor microenvironment and affecting tumor growth and transfer. OBJECTIVE:To explore the mechanisms of participation of bone marrow mesenchymal stem cel s in tumor tissue repair in an A549 lung cancer-bearing mouse model. METHODS:Bone marrow mesenchymal stem cel s were isolated in vitro, cultured, and identified using flow cytometry, and then used to establish a mouse model of A549 lung cancer-bearing. In the experimental group, human bone marrow mesenchymal stem cel s were injected into tissue surrounding the tumor. In the control group, an equal volume of PBS was injected. Animal survival condition and tumor size were compared. At 4 weeks, the specimens were harvested. Hematoxylin-eosin staining was used to compare tumor tissue. Masson staining was utilized to compare col agen fiber content. Reverse transcription-PCR was employed to detect the expression ofα-smooth muscle actin. Immunohistochemistry was used to examine the expression of fibroblast specific protein and fibroblast activation protein to reflect the degree of interstitial fibers in tumor tissue in both groups. The expression levels of vascular endothelial growth factor, hepatocyte growth factor, interleukin-6 and tenescin-C were compared between the two groups using immunohistochemistry. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cel s promoted tumor growth in tumor-bearing mice. The growth rate of tumor tissue in experimental group was faster than the control group (P<0.05). Compared with the control group,α-smooth muscle actin mRNA expression was significantly higher in the experimental group. Immunohistochemistry was used to detect the expression of tumor angiogenesis factors markers (fibroblast specific protein and fibroblast activation protein) in tumor tissue of experimental group. The expression levels of vascular endothelial growth factor, hepatocyte growth factor, interleukin-6 and tenescin-C were significantly greater in the experimental group than in the control group (P<0.05). Results indicated that bone marrow mesenchymal stem cel s differentiated into fibroblasts in tumor microenvironment, participated in the formation and construction of tumor stroma as wel as promoted the growth and repair of tumor via the secretion of vascular endothelial growth factor, hepatocyte growth factor, interleukin-6 and tenescin-C.
