[内源性神经节苷脂对神经母细胞瘤细胞整合素α2β1介导的与胶原黏附的影响]

Liu Zhi-Ping, Wen Fei-Qiu, Chen Yi-Xin, Wang Sha-Yan, Zhou Ke-Ying, Xia Quan

Department of Pediatrics, Second Clinical College of Medicine, Jinan University, Shenzhen, Guangdong 518020, China.

Zhongguo Dang Dai Er Ke Za Zhi. 2007 Feb;9(1):42-6.

OBJECTIVE

To study the effect of endogeneous gangliosides (Gls) on integrin alpha2beta1-mediated adhesion of neuroblastoma cells to collagen (Col).

METHODS

Neuroblastoma SK-N-SH cell line was cultured in the modified eagle's medium with the presence of 10 mum D-threo-1-phenyl-2-decanolamino-3-morphinolin-1-propanol (D-PDMP), an inhibitor of glucosylceramide synthase. Flow cytometry was used to detect the expression of integrin alpha2beta1 in the cell line. The effects of Mg2(+) and monoclonal antibodies to integrin alpha2beta1 on the adhesion of the cell line to immobilized Col were observed. The adhesion cell number was measured with the BCA method and presented with absorptance A570.

RESULTS

There was a high expression of integrin alpha2beta1 in the SK-N-SH cell line without D-PDMP treatment. Endogenous Gls in the cells were almost depleted after 6-day exposure to D-PDMP, but the integrin alpha2beta1 expression was not significantly changed. 1 mmoL/L Mg2(+) treatment increased significantly the number of adhesion cells in the SK-N-SH cell line. The adhesion to Col of the SK-N-SH cells exposed to D-PDMP which Gls was depleted was significantly reduced compared with the control SK-N-SH cells treated with 1 mmoL/L Mg2(+) (A570: 0.33 +/- 0.016 vs 0.57 +/- 0.033; P < 0.01). After endogeneous Gls was added into the Gls-depleted SK-N-SH cells, the adhesion of the cells was restored (A570: 0.52 +/- 0.035). The adhesion of SK-N-SH cells was significantly blocked by anti-alpha2 and anti-beta1 monoclonal antibodies, with A570 of 0.31 +/- 0.018 and 0.36 +/- 0.021 respectively.

CONCLUSIONS

Endogenous tumor Gls increases neuroblastoma cell adhesion to Col by regulating the function of integrin alpha2beta1, but has no effects on the integrin expression. It is suggested that tumor Gls may play a role in migration, invasion and metastasis of tumor cells.

目的

研究内源性神经节苷脂（Gls）对神经母细胞瘤细胞整合素α2β1介导的与胶原蛋白（Col）黏附的影响。

方法

将神经母细胞瘤SK-N-SH细胞系培养于含有10 μmol D-苏式-1-苯基-2-癸醇氨基-3-吗啉代-1-丙醇（D-PDMP）的改良伊格尔培养基中，D-PDMP为葡萄糖神经酰胺合酶抑制剂。采用流式细胞术检测该细胞系中整合素α2β1的表达。观察Mg2(+)和整合素α2β1单克隆抗体对该细胞系与固定化Col黏附的影响。用BCA法测定黏附细胞数，并以吸光度A570表示。

结果

未用D-PDMP处理的SK-N-SH细胞系中整合素α2β1高表达。经6天D-PDMP处理后，细胞内源性Gls几乎耗尽，但整合素α2β1表达无明显变化。1 mmol/L Mg2(+)处理显著增加了SK-N-SH细胞系中的黏附细胞数。与用1 mmol/L Mg2(+)处理的对照SK-N-SH细胞相比，经D-PDMP处理导致Gls耗尽的SK-N-SH细胞对Col的黏附显著降低（A570：0.33±0.016对0.57±0.033；P<0.01）。向Gls耗尽的SK-N-SH细胞中加入内源性Gls后，细胞的黏附得以恢复（A570：0.52±0.035）。抗α2和抗β1单克隆抗体显著阻断了SK-N-SH细胞的黏附，A570分别为0.31±0.018和0.36±0.021。