Li Yan-Hui, Sun Xiang, Wang Hong-Bo
Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
J Huazhong Univ Sci Technolog Med Sci. 2015 Jun;35(3):390-396. doi: 10.1007/s11596-015-1443-0. Epub 2015 Jun 14.
This study examined the effect of CD24 on anoikis of ovarian cancer cells. The expression of CD24 was detected by RT-PCR and Western blotting in ovarian cancer cells with high metastatic potential (HO-8910PM cells) and low metastatic potential (A2780 cells). Cell viability and cell proliferation were detected by MTT assay in suspension culture and adhesion culture. Soft agar culture was used to observe the colony formation. Anoikis was flow cytometrically detected. The results showed that the expression levels of CD24 mRNA and protein were significantly higher in HO-8910PM cells than in A2780 cells (P<0.01). In the suspension culture and soft agar culture, the HO-8910PM cells formed larger and more colonies (35.33 ± 5.51 vs. 16.67 ± 4.04; P<0.01), and showed a stronger resistance to anoikis than A2780 cells did (cell apoptosis rate: 5.93% ± 2 .38% vs. 16.32% ± 2.00%; P<0.01). After treated with CD24 monoclonal antibodies, the number of colony formed in HO-8910PM and A2780 cells was significantly decreased (9.33 ± 2.52 and 8.00 ± 2.00, respectively), and the anoikis rate of the two cell lines was also markedly increased (23.11% ± 2.87% and 28.36% ± 2.29%, respectively). Our study suggested that CD24 may play an important role in the development of anoikis resistance and CD24 can be used as a new therapeutic target to induce anoikis and inhibit metastasis in ovarian cancer.
本研究检测了CD24对卵巢癌细胞失巢凋亡的影响。采用RT-PCR和蛋白质免疫印迹法检测具有高转移潜能的卵巢癌细胞(HO-8910PM细胞)和低转移潜能的卵巢癌细胞(A2780细胞)中CD24的表达。通过MTT法检测悬浮培养和贴壁培养中的细胞活力和细胞增殖。采用软琼脂培养观察集落形成。通过流式细胞术检测失巢凋亡。结果显示,HO-8910PM细胞中CD24 mRNA和蛋白的表达水平显著高于A2780细胞(P<0.01)。在悬浮培养和软琼脂培养中,HO-8910PM细胞形成的集落更大、更多(35.33±5.51对16.67±4.04;P<0.01),并且与A2780细胞相比,对失巢凋亡的抗性更强(细胞凋亡率:5.93%±2.38%对16.32%±2.00%;P<0.01)。用CD24单克隆抗体处理后,HO-8910PM细胞和A2780细胞形成的集落数量显著减少(分别为9.33±2.52和8.00±2.00),并且这两种细胞系的失巢凋亡率也显著增加(分别为23.11%±2.87%和28.36%±2.29%)。我们的研究表明,CD24可能在失巢凋亡抗性的发展中起重要作用,并且CD24可作为诱导失巢凋亡和抑制卵巢癌转移的新治疗靶点。