CAJ | 학술논문

目的在细胞共培养微环境下,观察内皮素(ET)受体拮抗剂阻断内皮素轴对前列腺癌细胞PC3和成骨细胞SaOS_2相互作用的影响.方法利用细胞体外共培养系统,比较前列腺癌细胞PC3和成骨细胞SaOS_2在共培养微环境下和单独培养环境下细胞增殖的差异.共培养微环境下,分别以ET_A受体(ET_AR)拮抗剂BQ123和ET_B受体(ET_BR)拮抗剂BQ788阻断相应的ET受体,观察其对前列腺癌细胞和成骨细胞增殖的影响.同时,通过酶联免疫吸附试验(ELISA)法测定各组培养液中的ET-I浓度,通过逆转录.聚合酶链反应(RT-PCR)测定PC3细胞、SaOS_2:细胞ET-I及其受体的mRNA表达,比较它们的表达差异.结果与单独培养比较,共培养微环境下的PC3细胞和SaOS_2细胞的增殖数量均显著升高(P均＜0.05).ET_AR拮抗剂BQ123能阻断共培养微环境对SeOS:细胞的生长促进作用,部分阻断共培养微环境对PC3细胞的生长促进作用,而ETBR拮抗剂BQ788则无显著的干预作用.EUSA结果显示共培养组培养液中ET-1浓度[(14.26±1.06)βg/L/10~5个细胞]显著高于单独培养组[(6.58±0.74)pc/L/105个细胞,P＜0.05].RT-PCR结果显示PC3细胞表达ET-1和ET_AR,SaOS_2细胞只表达ET_AR;与单独培养比较,共培养微环境下PC3细胞ET-l的表达显著升高(P＜0.05),而PC3细胞和SaOS2细胞ET_AR的表达差异均无统计学意义(P＞0.05).结论 ET-1及ET_AR构成的内皮素轴是共培养微环境中前列腺癌细胞PC3和成骨细胞之间相互作用促进增殖的重要链接因子,而ETAR拮抗剂能有效地抑制两者间的相互作用,抑制PC3细胞增殖.
목적 재세포공배양미배경하,관찰내피소(ET)수체길항제조단내피소축대전렬선암세포PC3화성골세포SaOS_2상호작용적영향.방법 이용세포체외공배양계통,비교전렬선암세포PC3화성골세포SaOS_2재공배양미배경하화단독배양배경하세포증식적차이.공배양미배경하,분별이ET_A수체(ET_AR)길항제BQ123화ET_B수체(ET_BR)길항제BQ788조단상응적ET수체,관찰기대전렬선암세포화성골세포증식적영향.동시,통과매련면역흡부시험(ELISA)법측정각조배양액중적ET-I농도,통과역전록.취합매련반응(RT-PCR)측정PC3세포、SaOS_2:세포ET-I급기수체적mRNA표체,비교타문적표체차이.결과 여단독배양비교,공배양미배경하적PC3세포화SaOS_2세포적증식수량균현저승고(P균＜0.05).ET_AR길항제BQ123능조단공배양미배경대SeOS:세포적생장촉진작용,부분조단공배양미배경대PC3세포적생장촉진작용,이ETBR길항제BQ788칙무현저적간예작용.EUSA결과현시공배양조배양액중ET-1농도[(14.26±1.06)βg/L/10~5개세포]현저고우단독배양조[(6.58±0.74)pc/L/105개세포,P＜0.05].RT-PCR결과현시PC3세포표체ET-1화ET_AR,SaOS_2세포지표체ET_AR;여단독배양비교,공배양미배경하PC3세포ET-l적표체현저승고(P＜0.05),이PC3세포화SaOS2세포ET_AR적표체차이균무통계학의의(P＞0.05).결론 ET-1급ET_AR구성적내피소축시공배양미배경중전렬선암세포PC3화성골세포지간상호작용촉진증식적중요련접인자,이ETAR길항제능유효지억제량자간적상호작용,억제PC3세포증식.
Objective To block endothelin axis by endothelin(ET)receptor antagonist in the microenvironment of co-cultivation and observe its effects on interactions between prostate cancer cell PC3 and osteoblasts SaOS_2.Methods The interactions PC3 cells and SaOS_2 cells were observed by using an coculture experimental system in vitro.and proliferation of thesetwo cell lines between co-culture and culture alone was compared,respectively.ET-l receptor was blocked with selective ET_AR antagonist BQ-123 or ET_BR antagonist BQ-788 in the co-culture microenvironment,respectively,and its effect on proliferation of PC3 cells and SaOS_2 cells was observed.At the same time.ET-1 concentration of culture media was determined by enzyme-linked immuosorbent assay(ELISA),and mRNA expression of ET-1 and its receptor in PC3 cells and SaOS_2 cells was detected by reverse transcription-polymerase chain reaction(RT-PCR),and the expression differences of those between two kinds of culture rnieroenvironment were compared.Results The proliferation of PC3 cells and SaOS_2 cells in the co-culture microenvironment was significantly increased as compared with that in the culture alone microenvironment(P＜0.05).Selective ET_AR antagonist BQ-123 could completely block the promofive effect of co-culture microenvironment on growth of SaOS_2 cells,and partially block promotive effect of co-culture micreenvironment on growth of PC3 cell.Selective ET_BR antagonist BQ-788 had no significant blockade effect.The resuIt of ELISA showed that ET-1 Concentration of culture ,media in co-culture group[(14.26±1.06)βg/L/10~5'cells]Was significantly hisher than that in control group[(6.58±0.74)μg/10~5 cells],P＜0.05.RT-PCR revealed that PC3 cells expressed ET-l and ET_AR,and saos2 cells only expressed ET_AR.As Compared with control group,the ET-1 expression of PC3 cells in co-culture microenvironment was significantly higher(P＜0.05).and the ET_AR expression of PC3 cells or SaOS_2 cells in co-culture microenvironment had no statistically signiticant difference(P＞0.05).Conclision Ten endothelin axis composed of ET-1 and ETAR was the important link factor of interactions on promoting proliferation between PC3 cells and SaOS_2 cells in co-culture microenvironment.The ETAR antagonist could effectually inhibit the cellular interactions between them and inhibit the proliferation of PC3 cells.