岭南现代临床外科
嶺南現代臨床外科
령남현대림상외과
LINGNAN MODERN CLINICS IN SURGERY
2014年
2期
118-121
,共4页
刘成%毕良宽%许可慰%b
劉成%畢良寬%許可慰%b
류성%필량관%허가위%b
前列腺癌干细胞%CXCL12%CXC 趋化因子受体-4%多西紫杉醇%蛋白激酶B
前列腺癌榦細胞%CXCL12%CXC 趨化因子受體-4%多西紫杉醇%蛋白激酶B
전렬선암간세포%CXCL12%CXC 추화인자수체-4%다서자삼순%단백격매B
Prostate cancer stem cells%CXCL12%CXC chemokine receptor-4%Docetaxel%Protein kinase B (PKB/Akt)
目的观察前列腺癌干细胞中趋化因子受体-4（CXCR4）的表达，探索 CXCR4影响癌干细胞化疗敏感性的机制。方法流式细胞仪检测细胞表面 CXCR4阳性率；MTS 检测不同剂量多西紫杉醇对细胞活力的影响，绘制量效曲线，计算多西紫杉醇 IC50；分别使用 CXCL12或者抗CXCR4抗体激活或者阻断CXCR4后，检测多西紫杉醇化疗后前列腺癌干细胞细胞活力；蛋白免疫印迹法测 p-Akt、Akt 蛋白的表达变化。结果前列腺癌干细胞中 CXCR4阳性率为71．37±4．03％，非癌干为19．08±1．86％，二者有显著差异；使用多西紫杉醇化疗时，癌干细胞 IC50为7．5μM，非癌干细胞 IC50为0．6μM，癌干细胞耐多西紫杉醇能力明显强于非癌干细胞；CXCL12激活 CXCR4后，细胞活力为92．15±3．44％；抑制 CXCR4后，细胞活力为68．46±4．16％；对照组细胞活力为70．24±3．52％；使用 CXCL12激活 CXCR4后，Akt 磷酸化水平显著增加，抗 CXCR4抗体可以阻断Akt 磷酸化过程。结论 CXCL12/CXCR4可通过激活 Akt 下调前列腺癌干细胞对多西紫杉醇敏感性，这可能是前列腺癌干细胞化疗不敏感的重要机制之一。
目的觀察前列腺癌榦細胞中趨化因子受體-4（CXCR4）的錶達，探索 CXCR4影響癌榦細胞化療敏感性的機製。方法流式細胞儀檢測細胞錶麵 CXCR4暘性率；MTS 檢測不同劑量多西紫杉醇對細胞活力的影響，繪製量效麯線，計算多西紫杉醇 IC50；分彆使用 CXCL12或者抗CXCR4抗體激活或者阻斷CXCR4後，檢測多西紫杉醇化療後前列腺癌榦細胞細胞活力；蛋白免疫印跡法測 p-Akt、Akt 蛋白的錶達變化。結果前列腺癌榦細胞中 CXCR4暘性率為71．37±4．03％，非癌榦為19．08±1．86％，二者有顯著差異；使用多西紫杉醇化療時，癌榦細胞 IC50為7．5μM，非癌榦細胞 IC50為0．6μM，癌榦細胞耐多西紫杉醇能力明顯彊于非癌榦細胞；CXCL12激活 CXCR4後，細胞活力為92．15±3．44％；抑製 CXCR4後，細胞活力為68．46±4．16％；對照組細胞活力為70．24±3．52％；使用 CXCL12激活 CXCR4後，Akt 燐痠化水平顯著增加，抗 CXCR4抗體可以阻斷Akt 燐痠化過程。結論 CXCL12/CXCR4可通過激活 Akt 下調前列腺癌榦細胞對多西紫杉醇敏感性，這可能是前列腺癌榦細胞化療不敏感的重要機製之一。
목적관찰전렬선암간세포중추화인자수체-4（CXCR4）적표체，탐색 CXCR4영향암간세포화료민감성적궤제。방법류식세포의검측세포표면 CXCR4양성솔；MTS 검측불동제량다서자삼순대세포활력적영향，회제량효곡선，계산다서자삼순 IC50；분별사용 CXCL12혹자항CXCR4항체격활혹자조단CXCR4후，검측다서자삼순화료후전렬선암간세포세포활력；단백면역인적법측 p-Akt、Akt 단백적표체변화。결과전렬선암간세포중 CXCR4양성솔위71．37±4．03％，비암간위19．08±1．86％，이자유현저차이；사용다서자삼순화료시，암간세포 IC50위7．5μM，비암간세포 IC50위0．6μM，암간세포내다서자삼순능력명현강우비암간세포；CXCL12격활 CXCR4후，세포활력위92．15±3．44％；억제 CXCR4후，세포활력위68．46±4．16％；대조조세포활력위70．24±3．52％；사용 CXCL12격활 CXCR4후，Akt 린산화수평현저증가，항 CXCR4항체가이조단Akt 린산화과정。결론 CXCL12/CXCR4가통과격활 Akt 하조전렬선암간세포대다서자삼순민감성，저가능시전렬선암간세포화료불민감적중요궤제지일。
Objective To explore the expression of chemokine (C-X-C motif) receptor 4 (CXCR4) in prostate cancer stem cells (PCSCs), and the role of CXCR4 in PCSCs docetaxel resistance. Methods FACS was employed to measure the expression of CXCR4 on PCSCs and non-PCSCs surfaces; MTS was conducted to evaluate the cell viability under different concentration of docetaxel , and IC50 was calculated according to the dose-effect curves; CXCL12 or antibody against CXCR4 were used before docetaxel was added into the medium , and cell viability was measured by MTS; Western blotting was conducted to test the activation of signaling pathways. Results PCSCs cells contained 74.24% CXCR4 positive cells , while only 20.39% cells were CXCR4 positive in non-PCSCs (P<0.05); IC50 of docetaxelwas 7.5 μM in PCSCs, and 0.6 μM in non-PCSCs; 72 hours after docetaxel treatment, cell viability was 92.15±3.44% in CXCL12 group, 68.46±4.16% in anti-CXCR4 group, and 70.24±3.52% in negative control group; CXCL12 could significantly increase Akt phosphorylation in PCSCs. Conclusion CXCL12 activate the Akt signaling pathway via CXCR4 on PCSCs , inducing the docetaxel resistance of PCSCs , which may be one of the important mechanisms underlying the desensitization of docetaxel in PCSCs.
