84年鼠女哪年财运最旺,857comvvv色九欧美激情|85PO_87国产精品欲av国产av资源

[1]于雅麗,王雷鳴.二氯乙酸鈉對人骨肉瘤MG63細胞增殖、凋亡和遷移的影響[J].中醫(yī)正骨,2017,29(01):11-17.
　YU Yali,WANG Leiming.Effect of sodium dichloroacetate on cell proliferation,apoptosis and migration in human MG63 osteosarcoma cells[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(01):11-17.
點擊復(fù)制

二氯乙酸鈉對人骨肉瘤MG63細胞增殖、凋亡和遷移的影響()
分享到：

《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第29卷
期數(shù):
2017年01期
頁碼:
11-17
欄目:
基礎(chǔ)研究
出版日期:
2017-01-20

文章信息/Info

Title:
Effect of sodium dichloroacetate on cell proliferation,apoptosis and migration in human MG63 osteosarcoma cells
作者:
于雅麗; 王雷鳴
河南省鄭州市骨科醫(yī)院,河南 鄭州 450052
Author(s):
YU Yali; WANG Leiming
Zhengzhou Orthopedic Hospital,Zhengzhou 450052,Henan,China
關(guān)鍵詞:
骨肉瘤;  二氯乙酸鹽;  細胞增殖;  細胞凋亡;  細胞運動;  半胱氨酸天冬氨酸蛋白酶3;  MG63細胞株
Keywords:
osteosarcoma;  dichloroacetate;  cell proliferation;  apoptosis;  cell movement;  caspase 3;  MG63 cell lines
摘要:
目的:觀察二氯乙酸鈉(sodium dichloroacetate,DCA-Na)對人骨肉瘤MG63細胞增殖、凋亡和遷移的影響。方法:將對數(shù)生長期的MG63細胞隨機分為對照組和低、中、高濃度DCA-Na組,對照組不加DCA-Na,低、中、高DCA-Na組加入DCA-Na(終濃度分別為50 μg·mL-1、100 μg·mL-1、200 μg·mL-1),并設(shè)1個只加等量培養(yǎng)基不加細胞和DCA-Na的空白組。分別在干預(yù)24 h、48 h、72 h后,采用Cell Counting Kit-8細胞活性檢測試劑盒分光光度法檢測MG63細胞增殖情況,測定光密度(optical density,OD),計算低、中、高DCA-Na組細胞增殖抑制率,細胞增殖抑制率=[1-(DCA-Na組OD值-空白組OD值)/(對照組OD值-空白組OD值)]×100%; 分別采用Caspase-3活性檢測試劑盒分光光度法和Annexin V-FITC細胞凋亡檢測試劑盒流式細胞法檢測MG63細胞Caspase-3酶活性情況和細胞凋亡情況; 并在干預(yù)48 h后采用Transwell實驗檢測MG63細胞遷移情況。結(jié)果:①細胞增殖檢測結(jié)果。干預(yù)后,低、中、高DCA-Na組MG63細胞增殖抑制明顯,且隨干預(yù)時間的延長,3組細胞增殖抑制率均呈上升趨勢。各時間點間MG63細胞增殖抑制率差異有統(tǒng)計學(xué)意義(F=847.080,P=0.000),存在時間效應(yīng); 干預(yù)24 h、48 h、72 h后,低、中、高DCA-Na組MG63細胞增殖抑制率的組間差異均有統(tǒng)計學(xué)意義,且低DCA-Na組<中DCA-Na組<高DCA-Na組[(10.802±3.000)%,(18.792±2.261)%,(27.080±3.133)%,F=2.795,P=0.000;(13.098±1.299)%,(25.215±2.676)%,(44.382±2.397)%,F=4.362,P=0.000;(14.728±1.177)%,(35.297±4.757)%,(64.227±4.549)%,F=5.896,P=0.000]; 3組間MG63細胞增殖抑制率總體比較,差異有統(tǒng)計學(xué)意義,存在分組效應(yīng)(F=296.412,P=0.000); 時間因素與分組因素之間存在交互效應(yīng)(F=75.678,P=0.000)。②Caspase-3酶活性檢測結(jié)果。對照組MG63細胞Caspase-3酶活性無明顯變化,干預(yù)后低、中、高DCA-Na組MG63細胞Caspase-3酶活性均呈上升趨勢。各時間點間MG63細胞Caspase-3酶活性的差異有統(tǒng)計學(xué)意義(F=1 480.792,P=0.000),存在時間效應(yīng); 干預(yù)24 h、48 h、72 h后,4組間MG63細胞Caspase-3酶活性的差異均有統(tǒng)計學(xué)意義,且對照組<低DCA-Na組<中DCA-Na組<高DCA-Na組(0.027±0.003,0.143±0.005,0.153±0.008,0.161±0.003,F=2.320,P=0.000; 0.035±0.003,0.174±0.004,0.184±0.007,0.253±0.001,F=1.014,P=0.000; 0.031±0.004,0.246±0.006,0.275±0.003,0.371±0.004,F=1.000,P=0.000); 4組間MG63細胞Caspase-3酶活性總體比較,差異有統(tǒng)計學(xué)意義,存在分組效應(yīng)(F=624.975,P=0.000); 時間因素與分組因素之間存在交互效應(yīng)(F=94.579,P=0.000)。③流式細胞法細胞凋亡檢測結(jié)果。對照組MG63細胞凋亡率無明顯變化,干預(yù)后低、中、高濃度DCA-Na組MG63細胞凋亡率均呈上升趨勢。各時間點間MG63細胞凋亡率的差異有統(tǒng)計學(xué)意義(F=359.645,P=0.000),存在時間效應(yīng); 干預(yù)24 h、48 h、72 h后,各組間MG63細胞凋亡率的差異均有統(tǒng)計學(xué)意義,且對照組<低DCA-Na組<中DCA-Na組<高DCA-Na組[(2.554±0.427)%,(10.708±2.012)%,(20.857±2.531)%,(27.312±2.140)%,F=6.733,P=0.000;(1.748±0.202)%,(18.604±2.721)%,(29.471±1.605)%,(36.873±2.734)%,F=9.292,P=0.000;(1.944±0.112)%,(24.071±3.921)%,(30.050±3.921)%,(38.211±1.721)%,F=8.237,P=0.000]; 4組間MG63細胞凋亡率總體比較,差異有統(tǒng)計學(xué)意義,存在分組效應(yīng)(F=46.627,P=0.000); 時間因素與分組因素之間存在交互效應(yīng)(F=7.012,P=0.000)。④細胞遷移檢測結(jié)果。干預(yù)48 h后,4組遷移細胞數(shù)[顯微鏡每個視野下(×200)]的組間差異有統(tǒng)計學(xué)意義[(84.45±10.45)個,(74.56±9.45)個,(65.41±5.21)個,(40.21±4.52)個,F=148.243,P=0.000)]; 低、中、高DCA-Na組遷移細胞數(shù)均少于對照組(P=0.017,P=0.001,P=0.000),且低DCA-Na組>中DCA-Na組>高DCA-Na組(P=0.012,P=0.001,P=0.000)。結(jié)論:DCA-Na可抑制人骨肉瘤MG63細胞的增殖,增強MG63細胞Caspase-3酶活性,誘導(dǎo)和促進MG63細胞凋亡,抑制MG63細胞的遷移; 且濃度越高、干預(yù)時間越長,其影響越明顯。
Abstract:
Objective:To explore the effect of sodium dichloroacetate(DCA-Na)on cell proliferation,apoptosis and migration in human MG63 osteosarcoma cells.Methods:The MG63 cells were randomly divided into control group,low-concentration DCA-Na group,middle-concentration DCA-Na group and high-concentration DCA-Na group in the logarithmic growth phase.No DCA-Na were placed in cells in control group,and DCA-Na with final concentration of 50,100 and 200 μg/mL were placed in cells in low-,middle- and high-concentration DCA-Na group respectively,meanwhile,a blank group was established with same amount of culture medium and without cells and DCA-Na.At 24,48 and 72 hours after the intervention,the MG63 cell proliferations were detected by using Cell Counting Kit-8 spectrophotometry and the optical density(OD)were measured for calculating the cell proliferation inhibition rate([1-(ODDCA-Na group-ODblank group)/(ODcontrol group-ODblank group)]×100%)of low-,middle- and high-concentration DCA-Na group respectively.Meanwhile,The Caspase-3 enzymatic activity and apoptosis of MG63 cells were detected by using Caspase-3 activity assay kit spectrophotometry and Annexin V-FITC apoptosis assays kit flow cytometry respectively.The MG63 cell migration was detected by using Transwell assay at 48 hours after the intervention.Results:The MG63 cell proliferations were obviously inhibited after intervention in low-,middle- and high-concentration DCA-Na group,and a time-dependent rising trend of cell proliferation inhibition rate was found in the 3 groups.There was statistical difference in the MG63 cell proliferation inhibition rate between different timepoints(F=847.080,P=0.000),in other words,there was time effect.There was statistical difference in the MG63 cell proliferation inhibition rate between low-,middle- and high-concentration DCA-Na group at 24,48 and 72 hours after the intervention,and the MG63 cell proliferation inhibition rate was lower in the low-concentration DCA-Na group compared to middle-concentration DCA-Na group and was lower in the middle-concentration DCA-Na group compared to high-concentration DCA-Na group(10.802+/-3.000,18.792+/-2.261,27.080+/-3.133%,F=2.795,P=0.000; 13.098+/-1.299,25.215+/-2.676,44.382+/-2.397%,F=4.362,P=0.000; 14.728+/-1.177,35.297+/-4.757,64.227+/-4.549%,F=5.896,P=0.000).There was statistical difference in MG63 cell proliferation inhibition rate between the 3 groups in general,in other words,there was group effect(F=296.412,P=0.000).There was interaction between time factor and group factor(F=75.678,P=0.000).No significant change was found in Caspase-3 enzymatic activity of MG63 cells in control group,while a rising trend of the Caspase-3 enzymatic activity of MG63 cells was found in low-,middle- and high-concentration DCA-Na group after intervention.There was statistical difference in the Caspase-3 enzymatic activity of MG63 cells between different timepoints(F=1 480.792,P=0.000),in other words,there was time effect.There was statistical difference in the Caspase-3 enzymatic activity of MG63 cells between the 4 groups at 24,48 and 72 hours after the intervention,and the Caspase-3 enzymatic activity of MG63 cells presented a low-to-high trend in control group and low-,middle- and high-concentration DCA-Na group in turn(0.027+/-0.003,0.143+/-0.005,0.153+/-0.008,0.161+/-0.003,F=2.320,P=0.000; 0.035+/-0.003,0.174+/-0.004,0.184+/-0.007,0.253+/-0.001,F=1.014,P=0.000; 0.031+/-0.004,0.246+/-0.006,0.275+/-0.003,0.371+/-0.004,F=1.000,P=0.000).There was statistical difference in Caspase-3 enzymatic activity of MG63 cells between the 4 groups in general,in other words,there was group effect(F=624.975,P=0.000).There was interaction between time factor and group factor(F=94.579,P=0.000).No significant change was found in MG63 cell apoptosis rate in control group,while a rising trend of MG63 cell apoptosis rate was found in low-,middle- and high-concentration DCA-Na group after intervention.There was statistical difference in the MG63 cell apoptosis rate between different timepoints(F=359.645,P=0.000),in other words,there was time effect.There was statistical difference in the MG63 cell apoptosis rate between the 4 groups at 24,48 and 72 hours after the intervention,and the MG63 cell apoptosis rate presented a low-to-high trend in control group and low-,middle- and high-concentration DCA-Na group in turn(2.554+/-0.427,10.708+/-2.012,20.857+/-2.531,27.312+/-2.140%,F=6.733,P=0.000; 1.748+/-0.202,18.604+/-2.721,29.471+/-1.605,36.873+/-2.734%,F=9.292,P=0.000; 1.944+/-0.112,24.071+/-3.921,30.050+/-3.921,38.211+/-1.721%,F=8.237,P=0.000).There was statistical difference in the MG63 cell apoptosis rate between the 4 groups in general,in other words,there was group effect(F=46.627,P=0.000).There was interaction between time factor and group factor(F=7.012,P=0.000).There was statistical difference in the number of migratory MG63 cells under the optical microscope(×200)between the 4 groups at 48 hours after the intervention(84.45+/-10.45,74.56+/-9.45,65.41+/-5.21,40.21+/-4.52,F=148.243,P=0.000).The migratory MG63 cells was less in low-,middle- and high-concentration DCA-Na group compared to control group(P=0.017,P=0.001,P=0.000),and the low-concentration DCA-Na group surpassed middle- and high-concentration DCA-Na group and middle-concentration DCA-Na group surpassed high-concentration DCA-Na group in the number of migratory MG63 cells(P=0.012,P=0.001,P=0.000).Conclusion:DCA-Na can inhibit the proliferation of human MG63 osteosarcoma cells,increase the Caspase-3 enzymatic activity of MG63 cells,induce and accelerate the apoptosis of MG63 cells and inhibit the migration of MG63 cells.Moreover,the higher the concentration of DCA-Na is and the longer the intervention time is,the more obvious the effect is.

參考文獻/References:

[1] LI YC,ZHAO CH,YU ZB,et al.Low expression of miR-381 is a favorite prognosis factor and enhances the chemosensitivity of osteosarcoma[J].Oncotarget,2016,7(42):68585-68596.
[2] 郭會利,郭樹農(nóng),王軍輝,等.氯化鍶注射液治療轉(zhuǎn)移性骨腫瘤的臨床療效觀察[J].中醫(yī)正骨,2014,26(1):70-71.
[3] BOUVIER C,MACAGNO N,NGUYEN Q,et al.Prognostic value of the Hippo pathway transcriptional coactivators YAP/TAZ and beta 1-integrin in conventional osteosarcoma[J].Oncotarget,2016,7(40):64702-64710.
[4] ANDERSON ME.Update on survival in osteosarcoma[J].Orthop Clin North Am,2016,47(1):283-292.
[5] LETTIERI CK,APPEL N,LABBAN N,et al.Progress and opportunities for immune therapeutics in osteosarcoma[J].Immunotherapy,2016,8(10):1233-1244.
[6] 劉小軍.以異常能量代謝為靶點治療惡性腫瘤的研究進展[J].復(fù)旦學(xué)報:醫(yī)學(xué)版,2012,39(6):676-679.
[7] 菅記涌,張勇,朱大海.丙酮酸脫氫酶激酶的研究進展[J].基礎(chǔ)醫(yī)學(xué)與臨床,2008,28(11):1212-1215.
[8] WAKABAYASHI H,SAITO J,TAKI J,et al.Triple-phase contrast-enhanced MRI for the prediction of preoperative chemotherapeutic effect in patients with osteosarcoma:comparison with Tc-99m-MIBI scintigraphy[J].Skeletal Radiol,2016,45(1):87-95.
[9] MA Y,XU W,BAI R,et al.Volume-based predictive biomarkers of sequential FDG-PET/CT for sunitinib in cancer of unknown primary:identification of the best benefited patients[J].Eur J Nucl Med Mol Imaging,2016,44(2):199-205.
[10] 童晶濤.丙酮酸脫氫酶激酶-1在結(jié)腸癌中的表達及在細胞增殖和化療中的作用[D].重慶:第三軍醫(yī)大學(xué),2010.
[11] RICHARD SM,MARIGNAC VL.Sensitization to oxaliplatin in HCT116 and HT29 cell lines by metformin and ribavirin and differences in response to mitochondrial glutaminase inhibition[J].J Cancer Res Ther,2015,11(2):336-340.
[12] PARK SY,YOON JK,PARK KJ,et al.Prediction of occult lymph node metastasis using volume-based PET parameters in small-sized peripheralnon-small cell lung cancer[J].CancerImaging,2015:21.
[13] 朱明珍,茅國新,蔣華,等.二氯乙酸鈉誘導(dǎo)肺腺癌細胞株A549凋亡的實驗研究[J].現(xiàn)代醫(yī)藥衛(wèi)生,2014,30(3):321-323.
[14] LIU Z,SUN Y,TAN S,et al.Nutrient deprivation-related OXPHOS/glycolysis interconversion via HIF-1α/C-MYC pathway in U251 cells[J].Tumour Biol,2016,37(5):6661-6671.
[15] PRIGIONE A,ROHWER N,HOFFMANN S,et al.HIF1α modulates cell fate reprogramming through early glycolytic shift and upregulation of PDK1-3 and PKM2[J].Stem Cells,2014,32(2):364-376.

相似文獻/References:

[1]羅明星,劉丙根,羅文禮,等.保肢與截肢治療后骨肉瘤患者肢體功能及生活質(zhì)量的Meta分析[J].中醫(yī)正骨,2017,29(07):41.
　LUO Mingxing,LIU Binggen,LUO Wenli,et al.Comparison of limb salvage and amputation on limb function and life quality of patients with osteosarcoma:a meta analysis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(01):41.
[2]丁聚賢,謝興文,許偉,等.中醫(yī)藥在骨肉瘤放化療中的作用研究進展[J].中醫(yī)正骨,2018,30(05):43.
[3]羅漫麗,黃滿玉,李東升,等.多種血清腫瘤相關(guān)抗原自身抗體聯(lián)合檢測對骨肉瘤的診斷價值研究[J].中醫(yī)正骨,2021,33(04):32.
　LUO Manli,HUANG Manyu,LI Dongsheng,et al.A study of the clinical value of combined detection on serum autoantibodies against tumor-associated antigens in the diagnosis of osteosarcomas[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2021,33(01):32.
[4]劉柏良,秦瑨,劉小榮,等.miR-27a參與骨肉瘤發(fā)生發(fā)展作用機制的研究進展[J].中醫(yī)正骨,2025,37(03):53.

備注/Memo

備注/Memo:

通訊作者:王雷鳴 E-mial:[email protected]
更新日期/Last Update: 2017-01-20