生物化学与生物物理进展
生物化學與生物物理進展
생물화학여생물물리진전
PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
2009年
2期
238-243
,共6页
田辉%徐杰%刘贤锡%张冰%李文军%宋旭
田輝%徐傑%劉賢錫%張冰%李文軍%宋旭
전휘%서걸%류현석%장빙%리문군%송욱
鸟氨酸脱羧酶%S-甲硫氨酸脱羧酶%多胺%食管癌Eea109细胞%基因治疗
鳥氨痠脫羧酶%S-甲硫氨痠脫羧酶%多胺%食管癌Eea109細胞%基因治療
조안산탈최매%S-갑류안산탈최매%다알%식관암Eea109세포%기인치료
omithine decarboxylase%S-adenosylmethionine decarboxylase%polyamine%esophageal cancer Eeal09%gene therapy
为探讨ODC和AdoMetDC双反义腺病毒载体(Ad-ODC.AdoMetDCas)对食管癌Eca109细胞凋亡作用的影响,应用MTT法观察Ad-ODC-AdoMetDCas对食管癌Eca109细胞生长增殖的影响,采用Western blot和HPLC的方法分别检测腺病毒载体对食管癌Eea109细胞中ODC和AdoMetDC蛋白表达以及胞内多胺含量的抑制作用,同时应用原位未端标记(TUNEL)法观察Ad-ODC-AdoMetDCas对食管癌Eca109细胞凋亡作用的影响,透射电镜进一步观察细胞超微结构的改变.实验结果显示,应用MTT法观察发现Ad-ODC-AdoMetDCas对食管癌Eca109细胞生长增殖有显著抑制作用. 以Ad-ODC-AdoMetDCas感染食管癌Eca109细胞,可明显抑制食管癌Eca109细胞中ODC和AdoMetDC基因表达.HPLC结果显示,食管癌Eea109细胞感染Ad-ODC-AdoMetDCas后,细胞内3种多胺含量都明显降低.TUNEL标记检测结果显示Ad-ODC-AdoMetDCas可明显引起食管癌Eca109细胞凋亡.透射电镜观察到典型的细胞凋亡特征(表现细胞体积缩小,核皱缩、碎裂,染色质呈块状边集等).实验表明,ODC和AdoMetDC双反义腺病毒载体(Ad-ODC-AdoMetDCas)具有显著抑制食管癌细胞生长增殖,降低细胞多胺合成,促进细胞凋亡,为探讨食管癌基凶治疗的可行性提供实验依据.
為探討ODC和AdoMetDC雙反義腺病毒載體(Ad-ODC.AdoMetDCas)對食管癌Eca109細胞凋亡作用的影響,應用MTT法觀察Ad-ODC-AdoMetDCas對食管癌Eca109細胞生長增殖的影響,採用Western blot和HPLC的方法分彆檢測腺病毒載體對食管癌Eea109細胞中ODC和AdoMetDC蛋白錶達以及胞內多胺含量的抑製作用,同時應用原位未耑標記(TUNEL)法觀察Ad-ODC-AdoMetDCas對食管癌Eca109細胞凋亡作用的影響,透射電鏡進一步觀察細胞超微結構的改變.實驗結果顯示,應用MTT法觀察髮現Ad-ODC-AdoMetDCas對食管癌Eca109細胞生長增殖有顯著抑製作用. 以Ad-ODC-AdoMetDCas感染食管癌Eca109細胞,可明顯抑製食管癌Eca109細胞中ODC和AdoMetDC基因錶達.HPLC結果顯示,食管癌Eea109細胞感染Ad-ODC-AdoMetDCas後,細胞內3種多胺含量都明顯降低.TUNEL標記檢測結果顯示Ad-ODC-AdoMetDCas可明顯引起食管癌Eca109細胞凋亡.透射電鏡觀察到典型的細胞凋亡特徵(錶現細胞體積縮小,覈皺縮、碎裂,染色質呈塊狀邊集等).實驗錶明,ODC和AdoMetDC雙反義腺病毒載體(Ad-ODC-AdoMetDCas)具有顯著抑製食管癌細胞生長增殖,降低細胞多胺閤成,促進細胞凋亡,為探討食管癌基兇治療的可行性提供實驗依據.
위탐토ODC화AdoMetDC쌍반의선병독재체(Ad-ODC.AdoMetDCas)대식관암Eca109세포조망작용적영향,응용MTT법관찰Ad-ODC-AdoMetDCas대식관암Eca109세포생장증식적영향,채용Western blot화HPLC적방법분별검측선병독재체대식관암Eea109세포중ODC화AdoMetDC단백표체이급포내다알함량적억제작용,동시응용원위미단표기(TUNEL)법관찰Ad-ODC-AdoMetDCas대식관암Eca109세포조망작용적영향,투사전경진일보관찰세포초미결구적개변.실험결과현시,응용MTT법관찰발현Ad-ODC-AdoMetDCas대식관암Eca109세포생장증식유현저억제작용. 이Ad-ODC-AdoMetDCas감염식관암Eca109세포,가명현억제식관암Eca109세포중ODC화AdoMetDC기인표체.HPLC결과현시,식관암Eea109세포감염Ad-ODC-AdoMetDCas후,세포내3충다알함량도명현강저.TUNEL표기검측결과현시Ad-ODC-AdoMetDCas가명현인기식관암Eca109세포조망.투사전경관찰도전형적세포조망특정(표현세포체적축소,핵추축、쇄렬,염색질정괴상변집등).실험표명,ODC화AdoMetDC쌍반의선병독재체(Ad-ODC-AdoMetDCas)구유현저억제식관암세포생장증식,강저세포다알합성,촉진세포조망,위탐토식관암기흉치료적가행성제공실험의거.
Polyamine biosynthesis is controlled primarily by omithine Decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC). Antisense ODC and AdoMetDC sequences were cloned into an adenoviral vector (Ad-ODC-AdoMetDCas). To study the inhibitory effects of Ad-ODC-AdoMetDCas on polyamine biosynthesis and esophageal cancer cell apoptosis, adenovirus-mediated gene tmnsduction efficiency was assessed with counting GFP-positive cells using MTT. The malignant phenotype of Eca109 cells was assessed by growth curve. Western blot and HPLC were used to detect ODC and AdoMetDC expression and polyamine content in Ecal09 cells. TUNEL was used to analyze cell apoptosis. The change of morphology of apoptotic cells was observed by electron microscope. It was demonstrated approximate 70% of Eca 109 cells were infected with Ad-ODC-AdoMetDCas when MOI reached 50. The expression of ODC was inhibited in the infected tumor cells. Ad-ODC-AdoMetDCas could inhibit Ecal09 cell growth and invasive ability. TUNEL proved that Ad-ODC-AdoMetDCas can lead to cell apoptosis. Characterized morphology was observed by electron microscope (ehromatin condensation, nuclear disintegration, formation of apoptotic bodies). It was suggested Ad-ODC-AdoMetDCas has significant inhibitory effects on esophageal cancer cell proliferation, leads to cell apoptosis and bears therapeutic potential for the treatment of esophageal cancer.
