CAJ | 학술논문

目的研究白术挥发油对人前列腺癌细胞株的体外抗肿瘤作用. 方法常规培养LNCaP和DU145细胞株.设4个对照组:不含血清的空白培养液(A组),含血清的培养液(B组),含血清培养LNCaP细胞(C组),含血清培养DU145细胞(D组).设6个实验组,C组加入白术挥发油浓度分别50 μg/ml(C1组)、250 μg/ml(C2组)及500 μg/ml(C3组),D组分别加入上述3种浓度的白术挥发油(D1组、D2组、D3组).10组细胞铺24孔板,每组均设3个复孔,两种细胞均分别按每孔2×106铺板.常规培养48 h后,6个实验组加入各自浓度的白术挥发油,4个对照组以等量培养液替代.电镜观察各组细胞凋亡现象,取各组培养液上清并冷离心收集标本,分别检测睾酮(T)、雌二醇(E2)、孕酮(P)、血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(b-FGF)、tPSA和fPSA浓度.实验数据行多样本比较的统计学分析. 结果 C1和D2组生长抑制效应较好,LNCaP细胞呈时间效应正比关系,但DU145细胞仅在药物作用24 h后达到一次抑制率最大化(60.96％).与对照组相比,C、D组T浓度均为0,E2分别为269 pg/ml和239.81 pg/ml,呈高表达(P＜0.05)；P组间变化差异不显著；VEGF、b-FGF和fPSA均呈高表达,2组分别为102.96 pg/ml、0.26 ng/ml、0.16 ng/ml和1763.40 pg/ml、6.41 ng/ml、0.44 ng/ml,以D组更明显(P ＜0.05)；tPSA分别为0.36 ng/ml和0.发生凋亡的C1、C2、C3和D3组中,T最高由0升至0.37 ng/ml; E2最高由239.81 pg/ml升至649.90 pg/ml(P＜0.05)；P最高由0.98 ng/ml升至9.83 ng/ml(P＜ 0.01).VEGF、b-FGF和fPSA呈总体下降趋势.C2和D2组fPSA分别由0和0.04 ng/ml升至1.78 ng/ml和0.23 ng/ml. 结论白术挥发油对人前列腺癌细胞具有一定的凋亡诱导作用.雄激素非依赖性DU145细胞在性激素、细胞因子和PSA等表达方面与雄激素依赖性LNCaP细胞有不同的特点.
목적 연구백술휘발유대인전렬선암세포주적체외항종류작용. 방법 상규배양LNCaP화DU145세포주.설4개대조조:불함혈청적공백배양액(A조),함혈청적배양액(B조),함혈청배양LNCaP세포(C조),함혈청배양DU145세포(D조).설6개실험조,C조가입백술휘발유농도분별50 μg/ml(C1조)、250 μg/ml(C2조)급500 μg/ml(C3조),D조분별가입상술3충농도적백술휘발유(D1조、D2조、D3조).10조세포포24공판,매조균설3개복공,량충세포균분별안매공2×106포판.상규배양48 h후,6개실험조가입각자농도적백술휘발유,4개대조조이등량배양액체대.전경관찰각조세포조망현상,취각조배양액상청병랭리심수집표본,분별검측고동(T)、자이순(E2)、잉동(P)、혈관내피생장인자(VEGF)、감성성섬유세포생장인자(b-FGF)、tPSA화fPSA농도.실험수거행다양본비교적통계학분석. 결과 C1화D2조생장억제효응교호,LNCaP세포정시간효응정비관계,단DU145세포부재약물작용24 h후체도일차억제솔최대화(60.96％).여대조조상비,C、D조T농도균위0,E2분별위269 pg/ml화239.81 pg/ml,정고표체(P＜0.05)；P조간변화차이불현저；VEGF、b-FGF화fPSA균정고표체,2조분별위102.96 pg/ml、0.26 ng/ml、0.16 ng/ml화1763.40 pg/ml、6.41 ng/ml、0.44 ng/ml,이D조경명현(P ＜0.05)；tPSA분별위0.36 ng/ml화0.발생조망적C1、C2、C3화D3조중,T최고유0승지0.37 ng/ml; E2최고유239.81 pg/ml승지649.90 pg/ml(P＜0.05)；P최고유0.98 ng/ml승지9.83 ng/ml(P＜ 0.01).VEGF、b-FGF화fPSA정총체하강추세.C2화D2조fPSA분별유0화0.04 ng/ml승지1.78 ng/ml화0.23 ng/ml. 결론 백술휘발유대인전렬선암세포구유일정적조망유도작용.웅격소비의뢰성DU145세포재성격소、세포인자화PSA등표체방면여웅격소의뢰성LNCaP세포유불동적특점.
Objective To study the antitumor effects on human prostate cancer cell lines of a traditional Chinese medicine named atractylodes macrocephala koidz volatile oil in vitro.Methods LNCaP and DU145 cell lines were normally cultured and were divided into 4 control groups including:Blank culture without serum and without cell (group A),Blank culture with serum but without cell (group B),LNCaP cell culture with serum (group C),DU145 cell culture with serum (group D).In the meanwhile,there were 6 experimental groups:adding 50 μg/ml of atractylodes macrocephala koidz volatile oil in culture of group C1 and group D1,250ug/ml in group C2 and group D2,and 500ug/ml in group C3 and group D3.All of the 10 groups were simultaneously cultured in 24-well-plates for 48 hrs,and each group was repeatedly studied for three times.Forty-eight hours later,every 2 × 106 cells of LNCaP or DU145 was seeded into each well and atractylodes macrocephala koidz volatile oil was added in 6 experimental groups,and saline water was added into 4 control groups.Another 48 hrs later,the culture solutions of the 10 groups were separately collected for testosterone (T),estradiol (E2),progesterone (P),vascular endothelial growth factor (VEGF),basic fibroblast growth factor (b-FGF),total prostate specific antigen (tPSA) and free prostate specific antigen (fPSA) analysis with enzyme-linked immunoassay kits.The experiment was repeated for 3 times,and the mean data were statistically analyzed by SPSS One-way ANOVA.Results The growth of cultured cells was found to have been effectively inhibited in group C1 and group D2.The inhibition severity of LNCaP cells was positively related with the drug concentration and time,while DU145 cells could only be highly inhibited (60.96％) after 24hrs drug treatments.In group C and group D,we found that both T were in very low level (0 ng/ml) whereas both E2 were in high levels (269 pg/ml and 239.81 pg/ml,P ＜ 0.05),no distinct differences showed in P; In addition,VEGF,b-FGF and fPSA were all in high values,whose values were 102.96 pg/ml and 1763.40 pg/ml,0.26 ng/ml and 6.41 ng/ml,0.16 ng/ml and 0.44 ng/ml,respectively; DU145 cells had higher values than LNCaP cells (P ＜ 0.05).As regard to the 6 experimental groups,in the groups C1,C2,C3 and D3,we found that T had been unexpectedly increased from 0 to 0.37 ng/ml (P ＜ 0.05),E2 continuously elevated from 239.81 pg/ml to 649.90 pg/ml (P ＜0.05),surprisingly P were also increased from 0.98 ng/ml to 9.83 ng/ml (P ＜0.01).On the contrary,VEGF,b-FGF and fPSA levels were all graduallydecreased,dropping down to 47.79 pg/ml and 59.56pg/ml,0 and 1.79 ng/ml,0 and 0.11 ng/ml,respectively; nevertheless,in group C2 and group D2,fPSA values were surprisingly increased from 0 and 0.04 ng/ml up to 1.78 ng/ml and 0.23 ng/ml respectively.Conclusions Atractylodes macrocephala koidz volatile oil has certain anti-tumor effects on human prostate cancer.Compared with LNCaP cells,DU145 cells have many different characteristics in sex hormone,cytokine and PSA expressions.