中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2011年
6期
378-382
,共5页
李彦媚%高赛君%叶铁真%何映谊%林慧玲%朱欢欢
李彥媚%高賽君%葉鐵真%何映誼%林慧玲%硃歡歡
리언미%고새군%협철진%하영의%림혜령%주환환
基因,hermap%细胞分化%K562%细胞%信号转导通路
基因,hermap%細胞分化%K562%細胞%信號轉導通路
기인,hermap%세포분화%K562%세포%신호전도통로
Gene,hermap%Cell differentiation%K562 cells%Signal transduction pathway
目的 研究人类红细胞膜相关蛋白基因(hermap基因)对红细胞分化发育过程中信号转导通路关键激酶的影响,进一步揭示hermap基因对红细胞分化发育的作用.方法 分别建立稳定表达hermap 和 hermap-siRNA的K562细胞株,上调及下调K562细胞的hermap基因水平,并以阿糖胞苷(Ara-C)诱导K562细胞向红细胞分化发育,光学显微镜下观察诱导分化不同时间的细胞形态、联苯胺染色阳性细胞;FQ-PCR检测γ(Aγ、Gγ)珠蛋白基因和hermap基因表达量;流式细胞术(FCM)检测红细胞表面抗原CD36和CD235a,以及红细胞信号转导通路中关键激酶p-STATS、p-Akt、P-MAPK和p-c-JUN.结果 上调hermap表达后,随着培养时间延长,K562细胞体积、核质比例均逐渐减小,胞质由深蓝色逐渐转变为蓝紫色或浅红色,核染色质逐渐浓集、皱缩;联苯胺染色阳性细胞数(经Ara-C诱导24、48、72和96 h阳性细胞率分别为5.04%、20.42%、31.34%、48.88%,CD36+和CD235a+细胞数、γ(Aγ、Gγ)珠蛋白基因、hermap基因以及P-STAT5的表达量均逐渐增加;在培养的0、24、48、72、96 h,P-STAT5阳性细胞百分率分别为0.46%、4.54%、20.01%、23.65%、33.08%,与hermap基因表达量呈正相关(P<0.05).结论 hermap通过启动JAK/STAT5等一系列细胞转导信号,促进Ara-C诱导K562细胞向红细胞分化发育.
目的 研究人類紅細胞膜相關蛋白基因(hermap基因)對紅細胞分化髮育過程中信號轉導通路關鍵激酶的影響,進一步揭示hermap基因對紅細胞分化髮育的作用.方法 分彆建立穩定錶達hermap 和 hermap-siRNA的K562細胞株,上調及下調K562細胞的hermap基因水平,併以阿糖胞苷(Ara-C)誘導K562細胞嚮紅細胞分化髮育,光學顯微鏡下觀察誘導分化不同時間的細胞形態、聯苯胺染色暘性細胞;FQ-PCR檢測γ(Aγ、Gγ)珠蛋白基因和hermap基因錶達量;流式細胞術(FCM)檢測紅細胞錶麵抗原CD36和CD235a,以及紅細胞信號轉導通路中關鍵激酶p-STATS、p-Akt、P-MAPK和p-c-JUN.結果 上調hermap錶達後,隨著培養時間延長,K562細胞體積、覈質比例均逐漸減小,胞質由深藍色逐漸轉變為藍紫色或淺紅色,覈染色質逐漸濃集、皺縮;聯苯胺染色暘性細胞數(經Ara-C誘導24、48、72和96 h暘性細胞率分彆為5.04%、20.42%、31.34%、48.88%,CD36+和CD235a+細胞數、γ(Aγ、Gγ)珠蛋白基因、hermap基因以及P-STAT5的錶達量均逐漸增加;在培養的0、24、48、72、96 h,P-STAT5暘性細胞百分率分彆為0.46%、4.54%、20.01%、23.65%、33.08%,與hermap基因錶達量呈正相關(P<0.05).結論 hermap通過啟動JAK/STAT5等一繫列細胞轉導信號,促進Ara-C誘導K562細胞嚮紅細胞分化髮育.
목적 연구인류홍세포막상관단백기인(hermap기인)대홍세포분화발육과정중신호전도통로관건격매적영향,진일보게시hermap기인대홍세포분화발육적작용.방법 분별건립은정표체hermap 화 hermap-siRNA적K562세포주,상조급하조K562세포적hermap기인수평,병이아당포감(Ara-C)유도K562세포향홍세포분화발육,광학현미경하관찰유도분화불동시간적세포형태、련분알염색양성세포;FQ-PCR검측γ(Aγ、Gγ)주단백기인화hermap기인표체량;류식세포술(FCM)검측홍세포표면항원CD36화CD235a,이급홍세포신호전도통로중관건격매p-STATS、p-Akt、P-MAPK화p-c-JUN.결과 상조hermap표체후,수착배양시간연장,K562세포체적、핵질비례균축점감소,포질유심람색축점전변위람자색혹천홍색,핵염색질축점농집、추축;련분알염색양성세포수(경Ara-C유도24、48、72화96 h양성세포솔분별위5.04%、20.42%、31.34%、48.88%,CD36+화CD235a+세포수、γ(Aγ、Gγ)주단백기인、hermap기인이급P-STAT5적표체량균축점증가;재배양적0、24、48、72、96 h,P-STAT5양성세포백분솔분별위0.46%、4.54%、20.01%、23.65%、33.08%,여hermap기인표체량정정상관(P<0.05).결론 hermap통과계동JAK/STAT5등일계렬세포전도신호,촉진Ara-C유도K562세포향홍세포분화발육.
Objective To study the effects of hermap gene on kinases in erythroid signal transduction pathway and investigate the mechanism of hermap on erythroid differentiation. Methods The K562 cells expressing hermap and hermap-siRNA respectively were established for up- and down-regulating the expression of hermap gene. These K562 cells were then induced by Ara-C to erythroid differentiation and analyzed at 0, 24, 48 , 72 and 96 h, respectively, for cell morphology and biphenylamine staining positive cells, determination of CD235a, CD36, kinases p-STAT5, p-Akt, p-MAPK and p-c-JUN by FCM; and quantification of hermap gene and γ( Aγ,Gγ)globin gene by FQ-PCR. Results With up-regulating hermap gene and inducing by Ara-C, K562 cells were changing to low ratio of nucleus to cytoplasm, cytoplasm colour from basophilic to pinkish or amethyst tinge, increase of number of biphenylamine positive cells and expression of CD235a, CD36, γ( Aγ,Gγ)globin gene, hermap gene and p-STAT5 from 0 to 96 h. At 0, 24, 48, 72 and 96 h of culture, the positive rates of p-STAT5 cells were detected of 0. 46% , 4. 54% , 20. 01% , 23. 65% and 33.08% , respectively. This results demonstrated that there was a positive correlation between expression of p-STAT5 and hermap gene expression (P < 0.05). Conclusion hermap gene can stimulate erythroid differentiation of Ara-C induced K562 cells mainly through JAK/STAT5 signal transduction pathway.
