CAJ | 학술논문

目的：探讨人脐带间充质干细胞（hUCMSCs）在MCF－7B乳腺癌微环境中是否存在恶性转变并探究其生物学变化与STAT3异常激活及高表达的关系。方法将hUCMSCs分为3组：空白对照组（ hUCMSCs单独培养）、实验组（hUCMSCs与MCF－7B共培养）和阳性对照组（MCF－7B单独培养）。倒置显微镜观察细胞形态变化；流式细胞术检测细胞周期；RT－qPCR检测STAT3及其下游原癌基因c－Myc及抗凋亡Bcl－xLmRNA的表达；细胞免疫荧光检测p－STAT3、c－Myc和Bcl－xL蛋白表达及定位；Western blot检测p－STAT3、STAT3、c－Myc和Bcl－xL蛋白表达。结果实验组hUCMSCs与对照组比较核质比增大，细胞大小不一，排列紊乱；实验组细胞G1期比例显著低于对照组（ P＜0．05），S期和G2期比例均显著高于对照组（P＜0．05）；实验组STAT3、c－Myc和Bcl－xLmRNA的表达均显著高于对照组（P＜0．05）；实验组p－STAT3、STAT3、c－Myc和Bcl－xL的蛋白表达水平显著高于对照组（P＜0．05），且主要定位于细胞核内。结论 hUCMSCs在MCF－7B乳腺癌微环境中存在恶性转化趋势，且STAT3的异常激活及高表达是hUCMSCs恶性转化的重要因素之一。
목적：탐토인제대간충질간세포（hUCMSCs）재MCF－7B유선암미배경중시부존재악성전변병탐구기생물학변화여STAT3이상격활급고표체적관계。방법장hUCMSCs분위3조：공백대조조（ hUCMSCs단독배양）、실험조（hUCMSCs여MCF－7B공배양）화양성대조조（MCF－7B단독배양）。도치현미경관찰세포형태변화；류식세포술검측세포주기；RT－qPCR검측STAT3급기하유원암기인c－Myc급항조망Bcl－xLmRNA적표체；세포면역형광검측p－STAT3、c－Myc화Bcl－xL단백표체급정위；Western blot검측p－STAT3、STAT3、c－Myc화Bcl－xL단백표체。결과실험조hUCMSCs여대조조비교핵질비증대，세포대소불일，배렬문란；실험조세포G1기비례현저저우대조조（ P＜0．05），S기화G2기비례균현저고우대조조（P＜0．05）；실험조STAT3、c－Myc화Bcl－xLmRNA적표체균현저고우대조조（P＜0．05）；실험조p－STAT3、STAT3、c－Myc화Bcl－xL적단백표체수평현저고우대조조（P＜0．05），차주요정위우세포핵내。결론 hUCMSCs재MCF－7B유선암미배경중존재악성전화추세，차STAT3적이상격활급고표체시hUCMSCs악성전화적중요인소지일。
Objective_To investigate whether hUCMSCs undergo malignant transformation when exposed to MCF-7B breast cancer microenvironment and whether the abnormal activation and over expression of STAT3 play an impor-tant role in this transformation.Methods_The experiment was divided into three groups:blank group ( hUCMSCs were separately cultured),experimental group(hUCMSCs were indirectly co-cultured with MCF-7B breast cancer cells),positive control group(MCF-7B breast cancer cells were separately cultured).Morphology of cells was detec-ted by invertedmicroscope.Cell cycle was detected by flow cytometry.The mRNA expression of STAT3, c-Myc and Bcl-xL was tested by real-time PCR.The protein expression and location of p-STAT3,c-Myc and Bcl-xL were detected by immunofluorescence.The protein expressions of p-STAT3,STAT3,c-Myc and Bcl-xL were also meas-ured by Western blot.Results_The experimental group cells showed typical morphology of the tumor cells.The cells proportion of experiment group in G1 phase was significantly lower than that of the blank group(P<0.05),but which in S and G2 phase were significantly higher than those of the blank group(P<0.05).The mRNA expression levels of STAT3 ,c-Myc and Bcl-xL in experimental group was significantly higher than those in blank group ( P<0.05).p-STAT3,STAT3,c-Myc and Bcl-xL protein were significantly higher than those of the blank group(P<0.05),and they were mainly located in the nuclei.The protein expression of STATS also showed significant changes in experimental group.Conclusions_hUCMSCs trends to malignant transformations when exposed to MCF-7B breast cancer microenvironment.The abnormal activation and over expression of STAT3 are of important factors leading to the malignant transformation of hUCMSCs.