CAJ | 학술논문

目的探讨干扰CD147表达对白血病细胞系SHI-1细胞增殖、侵袭、转移能力的影响及其可能的机制.方法用慢病毒干扰载体及阴性对照载体转染SHI-1细胞,半定量RT-PCR法检测CD147、MMP-2及MMP-9 mRNA表达,Western blot法检测CD147蛋白水平;MTT法检测细胞增殖能力,用Millicell小室法将SHI-1细胞与骨髓基质细胞共培养检测SHI-1细胞体外侵袭能力的改变,将不同处理的SHI-1细胞经皮下或经尾静脉接种于裸鼠,观察SHI-1细胞在裸鼠体内增殖及浸润情况.结果慢病毒干扰载体转染的SHI-1细胞(SHI-1/CD147i细胞)CD147 mRNA表达水平较阴性对照病毒转染细胞(SHI-1/NC细胞)下调85％,CD147蛋白表达下降91％,SHI-1/CD147i细胞MMP-2及MMP-9mRNA表达较SHI-1/NC细胞分别下降72％及63％.SHI-1/CD 147i细胞体外增殖能力较SHI-1/NC及SHI-1细胞明显下降;与骨髓基质细胞共培养24 h后,SHI-1、SHI-1/NC、SHI-1/CD147i细胞移行至Millicell下层的细胞占接种细胞数的比例分别为(18.2±2.5)％、(16.5±2.7)％和(4.5±1.2)％,SHI-1/CD147i细胞移行能力显著低于SHI-1和SHI-1/NC细胞.接种于裸鼠皮下的SHI-1/CD147i细胞形成的肿瘤体积显著低于SHI-1及SHI-1/NC细胞.而经尾静脉接种SHI-1/CD147i细胞的裸鼠生存期较接种SHI-1、SHI-1/NC细胞的裸鼠显著延长,并且在脏器浸润能力下降.结论干扰SHI-1细胞CD147表达可抑制SHI-1细胞体内外增殖及浸润能力,其途径可能是通过下调MMP表达实现的.
목적 탐토간우CD147표체대백혈병세포계SHI-1세포증식、침습、전이능력적영향급기가능적궤제.방법 용만병독간우재체급음성대조재체전염SHI-1세포,반정량RT-PCR법검측CD147、MMP-2급MMP-9 mRNA표체,Western blot법검측CD147단백수평;MTT법검측세포증식능력,용Millicell소실법장SHI-1세포여골수기질세포공배양검측SHI-1세포체외침습능력적개변,장불동처리적SHI-1세포경피하혹경미정맥접충우라서,관찰SHI-1세포재라서체내증식급침윤정황.결과 만병독간우재체전염적SHI-1세포(SHI-1/CD147i세포)CD147 mRNA표체수평교음성대조병독전염세포(SHI-1/NC세포)하조85％,CD147단백표체하강91％,SHI-1/CD147i세포MMP-2급MMP-9mRNA표체교SHI-1/NC세포분별하강72％급63％.SHI-1/CD 147i세포체외증식능력교SHI-1/NC급SHI-1세포명현하강;여골수기질세포공배양24 h후,SHI-1、SHI-1/NC、SHI-1/CD147i세포이행지Millicell하층적세포점접충세포수적비례분별위(18.2±2.5)％、(16.5±2.7)％화(4.5±1.2)％,SHI-1/CD147i세포이행능력현저저우SHI-1화SHI-1/NC세포.접충우라서피하적SHI-1/CD147i세포형성적종류체적현저저우SHI-1급SHI-1/NC세포.이경미정맥접충SHI-1/CD147i세포적라서생존기교접충SHI-1、SHI-1/NC세포적라서현저연장,병차재장기침윤능력하강.결론 간우SHI-1세포CD147표체가억제SHI-1세포체내외증식급침윤능력,기도경가능시통과하조MMP표체실현적.
Objective To investigate the role of CD147 gene on the proliferation and infiltration of a human monocytic leukemic cell line SHI-1.Methods The expression of CD147 in SHI-1 cells was knockdowned by the lentiviral vector.The expressions of CD147,MMP-2 and MMP-9 were detected by semiquantitative RT-PCR.The protein of CD147 was detected by Western blotting.The capabilities of proliferation and infiltration of SHI-1 cell were examined by MTT and trans-matrigel invasion assay co-cultured with leukemia BMSC in vitro.SHI-1 cells were inoculated subcutaneously or via tail vein into nude mice to investigate its growth and infiltrative ability in vivo.Results The mRNA and protein of CD 147 in SHI-1/CD147i cells decreased by 85％ and 91％,respectively after the SHI-1 cells were infected by the lentivirus containing the CD147 siRNA.The proliferation capability of SHI-1/CD147i cells significantly decreased than those of SHI-1 and SHI-1/NC cells.The mRNA expressions of MMP-2、MMP-9 in SHI-1/CD147i cells were significantly lower than those in SHI-1/NC and SHI-1 cells.The SHI-1/CD147i cells showed significantly lower invasion rate than SHI-1 cells and SHI-1/NC cells when co-cultured with BMSCs.The neoplasms formed by SHI-1/CD147i cells in the subcutaneous of mice were significantly smaller than of the neoplasms formed by SHI-1 and SHI-1/NC cells.In nude mice inoculated via caudal vein with SHI/CD147i cells,mice demonstrated longer survival and moderate infiltration characteristic than those inoculated with SHI and SHI-1/NC cells.Conclusion CD147 might play important roles in the proliferation and infiltration of leukemia cells.CD147 should be a potential target for the treatment of acute leukemia.