目的:探討二氫丹參酮Ⅰ（Dihydrotanshinone I,DHI）對骨肉瘤細胞的抑制作用及其機制。方法:采用結晶紫染色法檢測濃度不同的DHI對人骨肉瘤細胞（MG-63、143B、Saos2、U2OS）增殖能力的影響;用MTT法進一步驗證DHI對人骨肉瘤細胞（MG-63、143B）增殖能力的作用。通過克隆形成實驗、Transwell遷移侵襲實驗、細胞劃痕實驗、Hoechst 33258染色以及應用流式細胞術等實驗方法檢測濃度不同濃度的DHI對人骨肉瘤細胞（MG-63、143B）克隆形成能力、遷移能力、侵襲能力及凋亡能力的影響。隨后用濃度不同的DHI（Control、0、2、4、6μM）處理人骨肉瘤143B細胞,通過蛋白印跡法（Western Blot）檢測用DHI處理的人骨肉瘤143B細胞和未用DHI處理的人骨肉瘤143B細胞中增殖細胞核抗原（PCNA）、Snail、MMP-2、MMP-7、MMP-9、N-Cadherin、Bcl-2、PARP、Cleaved–PARP、Caspase-3、Cleaved Caspase-3、LRP6、β-Catenin、Cyclin D1、c-M... 

【文章來源】：重慶醫(yī)科大學重慶市

【文章頁數(shù)】：46 頁

【學位級別】：碩士

【部分圖文】：

DHI抑制骨肉瘤細胞增殖Figure1DHIinhibitedOScellproliferationinvitro(A-F)DHIsuppressedOScellproliferation,asmeasuredbycrystalvioletstaining.(G-H)DHIsuppressedOScellproliferation,asmeasuredbytheMTTassay.(I-L)ColonyformationassaysshowedasignificantlylowercolonyformingrateinbothcelllinesfollowingDHItreatment.(M-N)

DHI抑制骨肉瘤細胞遷移和侵襲Figure2DHIinhibitedOScellsmigrationandinvasioninvitro(A–D)WoundhealingassaysshowedsignificantlydecreasedmigratoryabilitiesinbothcelllinesfollowingDHItreatment.(E–H)TranswellassaysshowedsignificantlydecreasedmigrationabilitiesinbothcelllinesfollowingDHItreatment.(I–L)TranswellassaysshowedsignificantlydecreasedinvasiveabilitiesinbothcelllinesfollowingDHItreatment.(M-N)Westernblotanalysisshowi

重慶醫(yī)科大學碩士研究生學位論文(A-D) Flow cytometry was used to explore whether DHI promoted OS cell apoptosis. (E-H)Hoechst staining showed that DHI significantly promoted apoptosis of two cell lines. (I-J) Westernblot analysis showing DHI downregulated Bcl-2, and upregulated PARP, Cleaved PARP, Caspase-3,Cleaved Caspase-3. (K) The Δψm (red/green) alteration in 143B cells was determined byfluorescence microscopy after staining with JC-1. μM: The different concentrations of DHI; Control:Osteosarcoma 143B cells were treated with DMSO to exclude the effect of DMSO on the test.△△P＜0.01, vs the control group; **P <0.01, vs 0μmol/L DHI group. (n=3)tu

【參考文獻】：
期刊論文
[1]二氫丹參酮Ⅰ對白血病THP1細胞的增殖抑制作用及其機制探討[J]. 李慧,何瑩,王慧潔,向洪,王春森.  實用醫(yī)院臨床雜志. 2018(02)
[2]表沒食子兒茶素沒食子酸酯對人骨肉瘤細胞143B的抑制作用及其機制探討[J]. 董超群,李方,徐麗,吉彩霞,劉曉驊,畢楊,羅進勇,李明.  腫瘤. 2017(06)
[3]Magnesium lithospermate B possesses inhibitory activity on Na+,K+-ATPase and neuroprotective effects against ischemic stroke[J]. Jason TC TZEN,Tzyy-rong JINN,Hank KH SHE,Balance CM CHEN,Vic HSIEH.  Acta Pharmacologica Sinica. 2007(05)



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