中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Traumatology
2014年
6期
621-625
,共5页
椎间盘%软骨%干细胞
椎間盤%軟骨%榦細胞
추간반%연골%간세포
Intervertebral disc%Cartilage%Stem cells
目的 探讨高表达HOXB4基因对人软骨终板干细胞(cartilage endplate stem cells,CESCs)增殖和细胞周期的影响及其意义. 方法 构建HOXB4基因腺病毒载体并转染至CESCs;将CESCs分为转染高表达HOXB4病毒组(A组),导入空病毒载体组(B组)和对照组.PCR及Western blot检测A组HOXB4基因及对应蛋白表达状况;采用细胞计数盒8(cell counting kit 8,CCK8)技术检测三组细胞增殖的差异,碘化丙啶(propidium iodide,PI)染色流式细胞仪检测三组细胞的细胞周期变化. 结果 (1)成功构建高表达HOXB4的病毒并顺利转染至CESCs;(2)实时定量PCR结果显示,转染病毒后CESCs的HOXB4的表达为对照组的3.6倍,Western blot 结果显示HOXB4蛋白表达为对照组的3倍;(3)HOXB4可促进CESCs的增殖(P＜0.05),并可将细胞阻断于S期,A组S期细胞从29.27升至30.28(P ＜0.05). 结论 高表达HOXB4可显著促进CESCs的增殖,并使细胞积聚于S期,可能是HOXB4延缓CESCs退变从而治疗腰椎间盘突出症的机制之一.
目的 探討高錶達HOXB4基因對人軟骨終闆榦細胞(cartilage endplate stem cells,CESCs)增殖和細胞週期的影響及其意義. 方法 構建HOXB4基因腺病毒載體併轉染至CESCs;將CESCs分為轉染高錶達HOXB4病毒組(A組),導入空病毒載體組(B組)和對照組.PCR及Western blot檢測A組HOXB4基因及對應蛋白錶達狀況;採用細胞計數盒8(cell counting kit 8,CCK8)技術檢測三組細胞增殖的差異,碘化丙啶(propidium iodide,PI)染色流式細胞儀檢測三組細胞的細胞週期變化. 結果 (1)成功構建高錶達HOXB4的病毒併順利轉染至CESCs;(2)實時定量PCR結果顯示,轉染病毒後CESCs的HOXB4的錶達為對照組的3.6倍,Western blot 結果顯示HOXB4蛋白錶達為對照組的3倍;(3)HOXB4可促進CESCs的增殖(P＜0.05),併可將細胞阻斷于S期,A組S期細胞從29.27升至30.28(P ＜0.05). 結論 高錶達HOXB4可顯著促進CESCs的增殖,併使細胞積聚于S期,可能是HOXB4延緩CESCs退變從而治療腰椎間盤突齣癥的機製之一.
목적 탐토고표체HOXB4기인대인연골종판간세포(cartilage endplate stem cells,CESCs)증식화세포주기적영향급기의의. 방법 구건HOXB4기인선병독재체병전염지CESCs;장CESCs분위전염고표체HOXB4병독조(A조),도입공병독재체조(B조)화대조조.PCR급Western blot검측A조HOXB4기인급대응단백표체상황;채용세포계수합8(cell counting kit 8,CCK8)기술검측삼조세포증식적차이,전화병정(propidium iodide,PI)염색류식세포의검측삼조세포적세포주기변화. 결과 (1)성공구건고표체HOXB4적병독병순리전염지CESCs;(2)실시정량PCR결과현시,전염병독후CESCs적HOXB4적표체위대조조적3.6배,Western blot 결과현시HOXB4단백표체위대조조적3배;(3)HOXB4가촉진CESCs적증식(P＜0.05),병가장세포조단우S기,A조S기세포종29.27승지30.28(P ＜0.05). 결론 고표체HOXB4가현저촉진CESCs적증식,병사세포적취우S기,가능시HOXB4연완CESCs퇴변종이치료요추간반돌출증적궤제지일.
Objective To observe the effect and significance of high-expression HOXB4 in controlling proliferation and cycle of human cartilage endplate stem cells (CESCs).Methods CESCs were divided into adenovirus-mediated HOXB4 delivery group (Group A),empty virus delivery group (Group B) and blank control group.Gene and protein expressions of HOXB4 in Group A were detected by PCR and Western blot respectively; cell proliferation among those groups were determined using cell counting kit 8 (CCK8) technique; cell cycle among those groups was measured by propidium iodide (PI) assay and flow cytometry.Results (1) Over-expressed HOXB4 virus was transferred to CESCs successfully; (2) Real-time quantitative PCR results showed 3.6 times higher expression of HOXB4 in Group A than in blank control group.Western blotting indicated HOXB4 protein in Group A was 3 times the level in control group; (3) HOXB4 promoted CESCs proliferation (P ＜ 0.05) and blocked the cells at phase S.Cells at phase S in Group A was increased from 29.27 to 30.28 (P ＜ 0.05).Conclusion Over-expressed HOXB4 accelerates proliferation of CESCs and increases cell population at phase S,indicating that HOXB4 hindering CESCs degeneration may be an approach to treat lumbar intervertebral disc protrusion.
