肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2013年
12期
809-811
,共3页
彭文岗%董胜利%樊晓龙%李阿奇
彭文崗%董勝利%樊曉龍%李阿奇
팽문강%동성리%번효룡%리아기
羟基磷灰石类%纳米粒%转染%结肠肿瘤%基因疗法
羥基燐灰石類%納米粒%轉染%結腸腫瘤%基因療法
간기린회석류%납미립%전염%결장종류%기인요법
Hydroxyapatites%Nanospheres%Transfection%Colonic neoplasms%Gene therapy
目的 探讨新型非病毒载体羟基磷灰石纳米粒子(nHAP)在肿瘤基因治疗中的作用及可能机制.方法 nHAP经氯化镁修饰后,经琼脂糖凝胶电泳判断nHAP-Mg2+与DNA的结合和保护作用.以增强型绿色荧光蛋白真核表达载体PEGFP-N1为报告基因,以脂质体作为基因载体转染到人类结肠癌SW480/M5细胞,流式细胞术测定转染效率和平均荧光值.四甲基偶氮唑蓝(MTT)比色法评价nHAP-Mg2+对细胞生长的影响.结果 在质量比合适时,nHAP-Mg2+与DNA能够完全结合并对DNA起到保护作用.单独应用nHAP-Mg2+时无基因转移功能,联合脂质体共同作为基因转染的载体可成功将PEGFP-N1基因导入SW480/M5细胞,其转染效率和平均荧光强度高于单用脂质体组(P＜0.05).单纯应用nHAP-Mg2+和联合脂质体时,随着nHAP-Mg2+量的增加(0 ～ 50 μg/ml)对SW480/M5细胞的生长抑制作用增强(P＜0.05).结论 nHAP-Mg2+具有结合和保护质粒DNA的作用.作为脂质体的辅助载体,不但可以提高基因转染效率和稳定表达,还可增强基因治疗的抗肿瘤效果.
目的 探討新型非病毒載體羥基燐灰石納米粒子(nHAP)在腫瘤基因治療中的作用及可能機製.方法 nHAP經氯化鎂脩飾後,經瓊脂糖凝膠電泳判斷nHAP-Mg2+與DNA的結閤和保護作用.以增彊型綠色熒光蛋白真覈錶達載體PEGFP-N1為報告基因,以脂質體作為基因載體轉染到人類結腸癌SW480/M5細胞,流式細胞術測定轉染效率和平均熒光值.四甲基偶氮唑藍(MTT)比色法評價nHAP-Mg2+對細胞生長的影響.結果 在質量比閤適時,nHAP-Mg2+與DNA能夠完全結閤併對DNA起到保護作用.單獨應用nHAP-Mg2+時無基因轉移功能,聯閤脂質體共同作為基因轉染的載體可成功將PEGFP-N1基因導入SW480/M5細胞,其轉染效率和平均熒光彊度高于單用脂質體組(P＜0.05).單純應用nHAP-Mg2+和聯閤脂質體時,隨著nHAP-Mg2+量的增加(0 ～ 50 μg/ml)對SW480/M5細胞的生長抑製作用增彊(P＜0.05).結論 nHAP-Mg2+具有結閤和保護質粒DNA的作用.作為脂質體的輔助載體,不但可以提高基因轉染效率和穩定錶達,還可增彊基因治療的抗腫瘤效果.
목적 탐토신형비병독재체간기린회석납미입자(nHAP)재종류기인치료중적작용급가능궤제.방법 nHAP경록화미수식후,경경지당응효전영판단nHAP-Mg2+여DNA적결합화보호작용.이증강형록색형광단백진핵표체재체PEGFP-N1위보고기인,이지질체작위기인재체전염도인류결장암SW480/M5세포,류식세포술측정전염효솔화평균형광치.사갑기우담서람(MTT)비색법평개nHAP-Mg2+대세포생장적영향.결과 재질량비합괄시,nHAP-Mg2+여DNA능구완전결합병대DNA기도보호작용.단독응용nHAP-Mg2+시무기인전이공능,연합지질체공동작위기인전염적재체가성공장PEGFP-N1기인도입SW480/M5세포,기전염효솔화평균형광강도고우단용지질체조(P＜0.05).단순응용nHAP-Mg2+화연합지질체시,수착nHAP-Mg2+량적증가(0 ～ 50 μg/ml)대SW480/M5세포적생장억제작용증강(P＜0.05).결론 nHAP-Mg2+구유결합화보호질립DNA적작용.작위지질체적보조재체,불단가이제고기인전염효솔화은정표체,환가증강기인치료적항종류효과.
Objective To investigate the role of hydroxyapatite nanoparticle (nHAP) in the gene transfection of human colorectal cancer cell line SW480/M5 and the possible mechanisms.Methods The combination and protection of nHAP-Mg2+ to DNA were analyzed by gelose gelatin electrophoresis.Liposome and nHAP modified by magnesium chloride was combined,and the PEGFP-N1 plasmids were transfected into SW480/M5 cells.The gene transfection rate and the mean fluorescence intensity were observed by flow cytometry.The effect of nHAP-Mg2+ on the growth of the cells were studied by MTT.Results At appropriate proportion,nHAP-Mg2+ could combine the plasmids compeletly and protected the DNA.The gene could not be transferred by nHAP-Mg2+ alone.Combining the nanoparticles and liposome,the gene could be transferred very efficiently and the transfection rates were significantly higher than the liposome (P ＜ 0.05).The inhibition of cell growth was increased along with the concentration of nHAP-Mg2+ wether it was used alone or with the combination of liposome (P ＜ 0.05).Conclusions nHAP-Mg2+ has the ability to combining and protecting DNA and can be used to transfer gene as the adjunct carrier of liposome for the gene therapy of tumor cells to elevate the gene tansfection and expression rate and also enhance the anti-tumor effection.
