中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2010年
5期
463-467
,共5页
任玉%周旋%原续波%许鹏%王广秀%贾志凡%张安玲%韩磊%浦佩玉%康春生
任玉%週鏇%原續波%許鵬%王廣秀%賈誌凡%張安玲%韓磊%浦珮玉%康春生
임옥%주선%원속파%허붕%왕엄수%가지범%장안령%한뢰%포패옥%강춘생
PAMAM%同载%As-miR-21%氟尿嘧啶%神经胶质瘤
PAMAM%同載%As-miR-21%氟尿嘧啶%神經膠質瘤
PAMAM%동재%As-miR-21%불뇨밀정%신경효질류
PAMAM%Co-delivery%As-miR-21%Fluorouracil%Glioma
目的 探讨as-miR-21增加U251脑胶质瘤细胞对5-FU化疗敏感性的效果.方法 透析法制备5-FU/PAMAM载体,透射电镜观察形态,紫外分光光度计检测载药率和包封率;流式细胞仪检测转染效率并分析细胞凋亡比例;MTT法检测共转染后细胞生长抑制效果;免疫荧光检测Ki67和Bcl-2蛋白表达;Trauswell检测细胞侵袭能力变化.结果 纳米微粒形态规整;平均包封率为66.21%,平均载药量为31.77%;PAMAM转染效率为70.53%;共转染组细胞生长显著抑制(F=273.345,P=0.000);凋亡比例升高(F=43.21,P=0.000),Ki67、Bcl-2表达均下调;细胞侵袭能力显著下降.结论 PAMAM可以有效同载as-miR-21和5-FU,且更加有效地抑制U251脑胶质瘤细胞的体外生长并增加对5-Fu的化疗敏感性.
目的 探討as-miR-21增加U251腦膠質瘤細胞對5-FU化療敏感性的效果.方法 透析法製備5-FU/PAMAM載體,透射電鏡觀察形態,紫外分光光度計檢測載藥率和包封率;流式細胞儀檢測轉染效率併分析細胞凋亡比例;MTT法檢測共轉染後細胞生長抑製效果;免疫熒光檢測Ki67和Bcl-2蛋白錶達;Trauswell檢測細胞侵襲能力變化.結果 納米微粒形態規整;平均包封率為66.21%,平均載藥量為31.77%;PAMAM轉染效率為70.53%;共轉染組細胞生長顯著抑製(F=273.345,P=0.000);凋亡比例升高(F=43.21,P=0.000),Ki67、Bcl-2錶達均下調;細胞侵襲能力顯著下降.結論 PAMAM可以有效同載as-miR-21和5-FU,且更加有效地抑製U251腦膠質瘤細胞的體外生長併增加對5-Fu的化療敏感性.
목적 탐토as-miR-21증가U251뇌효질류세포대5-FU화료민감성적효과.방법 투석법제비5-FU/PAMAM재체,투사전경관찰형태,자외분광광도계검측재약솔화포봉솔;류식세포의검측전염효솔병분석세포조망비례;MTT법검측공전염후세포생장억제효과;면역형광검측Ki67화Bcl-2단백표체;Trauswell검측세포침습능력변화.결과 납미미립형태규정;평균포봉솔위66.21%,평균재약량위31.77%;PAMAM전염효솔위70.53%;공전염조세포생장현저억제(F=273.345,P=0.000);조망비례승고(F=43.21,P=0.000),Ki67、Bcl-2표체균하조;세포침습능력현저하강.결론 PAMAM가이유효동재as-miR-21화5-FU,차경가유효지억제U251뇌효질류세포적체외생장병증가대5-Fu적화료민감성.
Objective To study the effect of as - miR - 21 enhance U251 human glioma cell chemo-sensitivity to 5 - FU. Method 5 - FU/PAMAM complex was prepared by dialysis method. As - miR - 21 was incubated with 5 - FU/PAMAM to form 5 - FU/PAMAM/as - miR - 21. Transmission electronic microscopy (TEM) was performed to observe the morphology of the nanoparticles. The drug loading efficiency and encapsulation efficiency was determined by ultraviolet spectroscopy ( UV ). The transfection of PAMAM dendrimer was detected by flow cytometry assay. MTT assay was carried out to determine U251 cell growth survival rate. Cell apoptosis was analyzed by flow - cytometry assay. Immunofluoresence staining was employed to determine Ki67 and Bcl -2 expression in U251 cells after combination treatment Transwell assay was performed to detect cell invasion ability. Results The morphology of the nanoparticle was sphere observed by TEM, and the diameter was less than 100 nm. The encapsulation efficiency and loading efficiency of drug were determined by ultraviolet spectroscopy to be 66. 21% and 31. 77% ,respectively. The results of flow cytometry assay showed that PAMAM dendrimer transfection efficiency was 70. 53%. The survival rate of U251 cells with as - miR - 21 and 5 - FU co - delivery treatment was significantly suppressed(F=273.345,P=0.000). The increased apoptosis nuclei percentage in co - delivery treated U251 cell was detected ( F =43. 21, P =0. 000) by flow - cytometry assay and the expression of Ki67 and Bcl -2 were down regulated simultaneously analyzed by immunofluorescence. Transwell result also demonstrated the cell invasion ability of U251 cells treated with 5 - FU combine with as - miR - 21 was decreased. Conclusions PAMAM dendrimer could effectively deliver as - miR - 21 and 5 - FU simultaneously;combination therapy can suppress U251 cell line growth effectively in vitro and can enhance the chemo -sensitivity of glioma cells to 5 - FU chemotherapy.