氯化鈷誘導(dǎo)低氧環(huán)境對(duì)人肝癌HepG2細(xì)胞株的生長(zhǎng)代謝影響.pdf

1、陳國(guó)風(fēng)：氯化鈷誘導(dǎo)低氧環(huán)境對(duì)人肝癌HepG2細(xì)胞株的生K4℃N影響氯化鈷誘導(dǎo)低氧環(huán)境對(duì)人肝癌HepG2細(xì)胞株的生長(zhǎng)代謝影響中文摘要1、目的建立人肝癌HepG2細(xì)胞在氯化鈷誘導(dǎo)的低氧環(huán)境下的生長(zhǎng)模型，觀察不同濃度氯化鈷誘導(dǎo)的低氧環(huán)境對(duì)人肝癌HepG2細(xì)胞的增殖、細(xì)胞的周期與凋亡即細(xì)胞的生長(zhǎng)狀況影響，進(jìn)而探討氯化鈷誘導(dǎo)的低氧對(duì)人肝癌HepG2細(xì)胞的生長(zhǎng)代謝的作用機(jī)制，并進(jìn)一步探討腫瘤細(xì)胞對(duì)低氧環(huán)境耐受性變化，為臨床上對(duì)腫瘤化療治療的療效尋找

2、理論依據(jù)。2、方法對(duì)數(shù)期人肝癌細(xì)胞株HepG2細(xì)胞，隨機(jī)分組對(duì)照組，不同濃度(50um／L、100um／L、150um／L、200IJm／L)Cocl：實(shí)驗(yàn)組，通過MTT法檢測(cè)HepG2細(xì)胞增殖情況計(jì)算細(xì)胞的抑制率并繪制HepG2細(xì)胞生長(zhǎng)抑制曲線；劃痕實(shí)驗(yàn)觀察HepG2細(xì)胞的遷移能力；通過流式細(xì)胞儀的Annexin—VFITC／PI雙染法和P工單染法檢測(cè)細(xì)胞的凋亡和周期變化；通過WesternBlot法檢測(cè)HepG2細(xì)胞的3一磷酸甘油醛

3、脫氫酶(Glyceraldehyde一3一phosphatedehydrogenase，GAPDH)、Bcl一2蛋白的表達(dá)。3、結(jié)果1通過MTT法檢測(cè)結(jié)果示在一定的時(shí)間和濃度范圍內(nèi)Cocl。抑制人肝癌HepG2細(xì)胞的增殖，并呈時(shí)間一劑量依賴性關(guān)系，即細(xì)胞的增值率隨著Cocl：濃度的增加和作用時(shí)間的延長(zhǎng)逐漸減低。2HepG2細(xì)胞經(jīng)過劃痕損傷試驗(yàn)后，實(shí)驗(yàn)組的細(xì)胞的遷移能力顯著變慢，對(duì)照組在經(jīng)過劃痕實(shí)驗(yàn)24h后，細(xì)胞已基本遷移至劃痕的中央，而

4、實(shí)驗(yàn)組隨著氯化鈷濃度的增加，細(xì)胞劃痕損傷面積未見明顯減少，劃痕明顯，說明氯化鈷抑制HepG2細(xì)胞的遷移及損傷修復(fù)能力，呈濃度依賴性關(guān)系。3流式細(xì)胞儀的檢測(cè)結(jié)果顯示：對(duì)照組、不同濃度(50um／L、100um／L、200um／L、300um／L、500lim／L)的實(shí)驗(yàn)組的細(xì)胞凋亡率(％)分別是：342、774、1307、2056、2853、4445(PO05)，與對(duì)照組相比較試驗(yàn)組的凋亡率明顯增加，并呈濃度依賴性。細(xì)胞的周期結(jié)果顯示實(shí)驗(yàn)

5、組的HepG2細(xì)胞隨著氯化鈷濃度的增加，處于細(xì)胞周期G。期的比例(％)由陳國(guó)風(fēng)：氯化鈷誘導(dǎo)低氧環(huán)境對(duì)人肝癌HepG2細(xì)胞株的生長(zhǎng)代謝影響TheeffectsofhypoxiainducedbythecobaltchlorideonthegrowthandapoptosisinHepaticCancerCellHepG2ObjeetiveAbstractToestablishthegrowthmodelofhumanhepatocellu

6、larcarcinomaHepG2cellsofcobaltchloride，observethehypoxiaenvironmentinducedbycobaltchlorideonhumanhepatocellularcarcinomaHepG2cellsproliferation，cellcycleandapoptosis，thendiscussesthemetabolismmechanismofhumanhepatocellul

7、arcarcinomaHepG2cellsofthehypoxiainducedbydifferentconcentrationsofcobaltchlorideonthegrowth，andthenfurthertoexplorethetolerancechangesofthetumorcellstothelowoxygenenvironmentwhichishelpfultofindthetheoreticalfoundationf

8、ortheclinicallytreatmentoftumorchemotherapyMethodLogarithmicphaseofHepG2celllinewererandomdividedintogroupscontrolgroupandexperimentalgroupwhichhadfivedifferentconcentrations(50prn／L、1009m／L，2009m／L、3009m／L、5009m／L)。MTTm

9、ethodassaywasusedtodetectcellsurvivalrateorcellproliferationcapacity，ScratchtestwereusedtoobservetheinfluenceofcobaltchloridewithdifferentdoesonmigrationabilityofHepG2celllineByflowcytometryofVFITC／PIdoublestainingmethod

10、andPIsinglestainingmethodtodetectcellapoptosisandcyclesrespectively，theexpressionsofGlyceraldehyde3phosphoricaciddehydrogenaseandtheBcl2proteinwereanalyzedbyWestemblotResult1BytheresultsofMTTmethodassaytheCobaltchloridei

11、nhibittheproliferationofhumanhepatocellularcarcinomaHepG2cells，andtheinhibitionwasontimedosedependentrelationshipinacertainreactingtimeandconcentrtions，namelythecellincrementratewasgraduallyreducedwiththeincreaseofconcen

12、trationsandtimeoftheCobaltchloride2TheeffectsoftheHepG2cellswasinjuredafterScratchtestwiththeincreasedconcentrationofcobaltchloridetheexperimentlgroupcellmigrationabilitywasrestrainedobviouslycomparedwithcontrolgroup，cob