西安交通大学学报（医学版）
西安交通大學學報（醫學版）
서안교통대학학보（의학판）
Journal of Xi'an Jiaotong University (Medical Sciences)
2015年
6期
775-781
,共7页
赵瑞华%李向柯%姜文静%张伟杰%宗红
趙瑞華%李嚮柯%薑文靜%張偉傑%宗紅
조서화%리향가%강문정%장위걸%종홍
CCL18%乳腺癌%miR98%miRNA%c-myc%N-Ras%lin28
CCL18%乳腺癌%miR98%miRNA%c-myc%N-Ras%lin28
CCL18%유선암%miR98%miRNA%c-myc%N-Ras%lin28
CCL18%breast cancer%miR98%miRNA%c-myc%N-Ras%lin28
目的：探讨 CCL18是否参与了乳腺癌 miRNAs 的表达调控。方法采用 miRNAs 芯片筛查 CCL18处理前后乳腺癌细胞 miRNAs 的表达差异,QRT-PCR 和 Luciferase Reporter Assay 对芯片结果进行验证。瞬时转染法分别改变乳腺癌细胞 miR98和 c-myc 的表达,应用 QRT-PCR 和 Western blot 检测 c-myc 和 lin28 mRNA 和蛋白的表达。结果miRNAs 芯片结果显示 CCL18处理后乳腺癌细胞共20种 miRNAs 发生变化,QRT-PCR 和 Luciferase Report-er Assay 证实 CCL18下调乳腺癌细胞 miR98的表达。CCL18可上调乳腺癌细胞 c-myc 和 lin28 mRNA 和蛋白的表达,转染 c-myc siRNAs 可逆转 CCL18调控 lin28和 miR98表达的功能。CCL18通过 miR98转录后调控乳腺癌细胞N-Ras 蛋白的表达。结论CCL18通过 N-Ras/c-myc/lin28通路下调 miR98,下调的 miR98通过 转 录 后 调 控 增 加N-Ras蛋白的表达,从而进一步激活 c-myc/lin28通路,维持 miR98的持续降低,形成了一个正反馈环路。
目的：探討 CCL18是否參與瞭乳腺癌 miRNAs 的錶達調控。方法採用 miRNAs 芯片篩查 CCL18處理前後乳腺癌細胞 miRNAs 的錶達差異,QRT-PCR 和 Luciferase Reporter Assay 對芯片結果進行驗證。瞬時轉染法分彆改變乳腺癌細胞 miR98和 c-myc 的錶達,應用 QRT-PCR 和 Western blot 檢測 c-myc 和 lin28 mRNA 和蛋白的錶達。結果miRNAs 芯片結果顯示 CCL18處理後乳腺癌細胞共20種 miRNAs 髮生變化,QRT-PCR 和 Luciferase Report-er Assay 證實 CCL18下調乳腺癌細胞 miR98的錶達。CCL18可上調乳腺癌細胞 c-myc 和 lin28 mRNA 和蛋白的錶達,轉染 c-myc siRNAs 可逆轉 CCL18調控 lin28和 miR98錶達的功能。CCL18通過 miR98轉錄後調控乳腺癌細胞N-Ras 蛋白的錶達。結論CCL18通過 N-Ras/c-myc/lin28通路下調 miR98,下調的 miR98通過 轉 錄 後 調 控 增 加N-Ras蛋白的錶達,從而進一步激活 c-myc/lin28通路,維持 miR98的持續降低,形成瞭一箇正反饋環路。
목적：탐토 CCL18시부삼여료유선암 miRNAs 적표체조공。방법채용 miRNAs 심편사사 CCL18처리전후유선암세포 miRNAs 적표체차이,QRT-PCR 화 Luciferase Reporter Assay 대심편결과진행험증。순시전염법분별개변유선암세포 miR98화 c-myc 적표체,응용 QRT-PCR 화 Western blot 검측 c-myc 화 lin28 mRNA 화단백적표체。결과miRNAs 심편결과현시 CCL18처리후유선암세포공20충 miRNAs 발생변화,QRT-PCR 화 Luciferase Report-er Assay 증실 CCL18하조유선암세포 miR98적표체。CCL18가상조유선암세포 c-myc 화 lin28 mRNA 화단백적표체,전염 c-myc siRNAs 가역전 CCL18조공 lin28화 miR98표체적공능。CCL18통과 miR98전록후조공유선암세포N-Ras 단백적표체。결론CCL18통과 N-Ras/c-myc/lin28통로하조 miR98,하조적 miR98통과 전 록 후 조 공 증 가N-Ras단백적표체,종이진일보격활 c-myc/lin28통로,유지 miR98적지속강저,형성료일개정반궤배로。
Objective To explore whether CCL18 is involved in regulating the expression of miRNAs in breast cancer.Methods The expression profile of miRNAs in the breast cancer cell following CCL18 treatment was determined by miRNAs microarray analysis.Then we performed QRT-PCR and Luciferase Reporter Assay to validate the results from the miRNAs microassay.We used transient transfection to change the expression of miR98 and c-myc in breast cancer cells.We then used QRT-PCR and Western blot to analyze the mechanism by which CCL18 downregulates the expression of miR98 in breast cancer cells.Results miRNAs microarray analysis showed that cells treated with CCL18 differentially expressed 20 miRNAs genes compared with those in the control group. Our QRT-PCR and Luciferase Reporter Assay confirmed the result.The mRNA and protein expressions of C-myc and lin28 were increased after CCL18 stimulation in breast cancer cells.Transfection with c-myc siRNAs rescued the increase of lin28 and loss of miR98 expression caused by CCL18 stimulation.Our results also showed that CCL18 could upregulate the expression of N-Ras at post-transcription level.Conclusion CCL18 downregulates the expression of miR98 via N-Ras/c-myc/lin28 pathway.The downregulated miR98 increases the expression of N-Ras after transfection,which further activates c-myc/lin28 pathway and forms a positive feedback loop.
