Tu Yan, Li Zhenjiang, Tang Aiping, Fei Yan, Li Huihui, Wu Qiong, He Wenfeng
Department of Hematology, the Second Hospital Affiliated to Nanchang University, Nanchang 330006, China.
Zhonghua Xue Ye Xue Za Zhi. 2014 Aug;35(8):737-42. doi: 10.3760/cma.j.issn.0253-2727.2014.08.015.
To investigate the role of CD147 gene on the proliferation and infiltration of a human monocytic leukemic cell line SHI-1.
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.
The mRNA and protein of CD147 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.
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.
探讨CD147基因对人单核细胞白血病细胞系SHI-1增殖和浸润的作用。
采用慢病毒载体敲低SHI-1细胞中CD147的表达。通过半定量RT-PCR检测CD147、基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）的表达。通过蛋白质印迹法检测CD147蛋白。采用MTT法和与白血病骨髓间充质干细胞（BMSC）共培养的基质胶侵袭试验检测SHI-1细胞的增殖和浸润能力。将SHI-1细胞皮下接种或经尾静脉接种到裸鼠体内，观察其在体内的生长和浸润能力。
含CD147小干扰RNA（siRNA）的慢病毒感染SHI-1细胞后，SHI-1/CD147i细胞中CD147的mRNA和蛋白分别下降了85%和91%。SHI-1/CD147i细胞的增殖能力明显低于SHI-1和SHI-1/阴性对照（NC）细胞。SHI-1/CD147i细胞中MMP-2、MMP-9的mRNA表达明显低于SHI-1/NC和SHI-1细胞。与BMSCs共培养时，SHI-1/CD147i细胞的侵袭率明显低于SHI-1细胞和SHI-1/NC细胞。SHI-1/CD147i细胞在小鼠皮下形成的肿瘤明显小于SHI-1和SHI-1/NC细胞形成的肿瘤。经尾静脉接种SHI/CD147i细胞的裸鼠比接种SHI和SHI-1/NC细胞的裸鼠生存期更长，浸润特征更轻。
CD147可能在白血病细胞的增殖和浸润中起重要作用。CD147应是急性白血病治疗的潜在靶点。
