Zheng Ying, Wang Xuemei, Wang Haidong, Yan Wei, Zhang Quan, Chang Xin
Department of Anesthesia, Huai'an First People's Hospital, Nanjing Medical University, 6 Beijing Road West, Huai'an, Jiangsu, 223300, People's Republic of China,
Tumour Biol. 2014 Jun;35(6):5189-98. doi: 10.1007/s13277-014-1673-y. Epub 2014 Feb 27.
Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. Previous studies have suggested that abnormal expression of BMP-4, BMP-7, and BMP-9 is correlated with tumor progression in HCC, but the role played by BMP-2 in HCC has not yet been reported. To determine the role of BMP-2 in HCC, we first investigated the effect of exogenous BMP-2 on the growth of the cell lines HCC SK-Hep-1, Hep G2, and Hep 3B. Next, we studied the function of BMP-2 in SK-Hep-1 HCC cell line using a recombinant lentivirus vector to deliver BMP-2. We also used siRNA to silence endogenous BMP-2 expression in the HCC Hep 3B cell line. Then, cell growth and migration were assayed in vitro using WST-8, wound-healing, and transwell invasion assays. Cellular apoptosis and cell-cycle distribution were assessed using flow cytometry. We also investigated the effects of BMP-2 overexpression and knockdown on the expression of proliferating cell nuclear antigen (PCNA), matrix metallopeptidase-2 (MMP-2), phosphorylated AKT (p-AKT), phosphoinositide 3-kinase p85α (PI3Kp85α), Bax, Bcl-2, caspase-3, cleaved caspase-3, p21, and cyclin E. As a result, we observed that BMP-2 inhibited the proliferation of HCC cells. Furthermore, HCC cell proliferation and migration were significantly diminished by BMP-2 overexpression, as was indicated by WST-8, would healing, and transwell assays, while knockdown of BMP-2 led to an increase in proliferation and migration of Hep 3B cells. BMP-2 overexpression significantly increased the susceptibility of SK-Hep-1 cells to low-serum-induced apoptosis, while BMP-2 knockdown reduced the susceptibility of Hep 3B cells. Overexpression of BMP-2 induced G1 phase arrest through upregulation of p21. When BMP-2 expression was elevated in SK-Hep-1 cells, the expression of PI3Kp85α, p-AKT, PCNA, and MMP-2 declined. These results suggest that BMP-2 exerts an inhibitory effect on the growth and migration of HCC cells, possibly via a blockade of PI3K/AKT signaling.
肝细胞癌（HCC）是全球第五大常见癌症。先前的研究表明，骨形态发生蛋白-4（BMP-4）、骨形态发生蛋白-7（BMP-7）和骨形态发生蛋白-9（BMP-9）的异常表达与HCC的肿瘤进展相关，但BMP-2在HCC中的作用尚未见报道。为了确定BMP-2在HCC中的作用，我们首先研究了外源性BMP-2对肝癌细胞系SK-Hep-1、Hep G2和Hep 3B生长的影响。接下来，我们使用重组慢病毒载体递送BMP-2，研究了BMP-2在SK-Hep-1肝癌细胞系中的功能。我们还使用小干扰RNA（siRNA）沉默肝癌Hep 3B细胞系中的内源性BMP-2表达。然后，使用WST-8、伤口愈合和Transwell侵袭试验在体外检测细胞生长和迁移。使用流式细胞术评估细胞凋亡和细胞周期分布。我们还研究了BMP-2过表达和敲低对增殖细胞核抗原（PCNA）、基质金属蛋白酶-2（MMP-2）、磷酸化AKT（p-AKT）、磷酸肌醇3激酶p85α（PI3Kp85α）、Bax、Bcl-2、半胱天冬酶-3、裂解的半胱天冬酶-3、p21和细胞周期蛋白E表达的影响。结果，我们观察到BMP-2抑制肝癌细胞的增殖。此外，WST-8、伤口愈合和Transwell试验表明，BMP-2过表达显著降低了肝癌细胞的增殖和迁移，而敲低BMP-2导致Hep 3B细胞的增殖和迁移增加。BMP-2过表达显著增加了SK-Hep-1细胞对低血清诱导凋亡的敏感性，而BMP-2敲低降低了Hep 3B细胞的敏感性。BMP-2过表达通过上调p21诱导G1期阻滞。当SK-Hep-1细胞中BMP-2表达升高时，PI3Kp85α、p-AKT、PCNA和MMP-2的表达下降。这些结果表明，BMP-2可能通过阻断PI3K/AKT信号传导对肝癌细胞的生长和迁移发挥抑制作用。
