Knockdown of biglycan expression by RNA interference inhibits the proliferation and invasion of, and induces apoptosis in, the HCT116 colon cancer cell line
Knockdown of biglycan expression by RNA interference inhibits the proliferation and invasion of, and induces apoptosis in, the HCT116 colon cancer cell line
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DOI：
10.3892/mmr.2015.4383
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发表时间：
2015-11-01
影响因子：
3.4
通讯作者：
Ma, Tianfei
中科院分区：
文献类型：
--
作者：
Xing, Xiaojing;Gu, Xiaohu;Ma, Tianfei
双糖蛋白聚糖是细胞外基质的重要组成部分，也是富含亮氨酸的小分子蛋白聚糖家族的一员。先前的研究表明，双糖链蛋白聚糖的表达在多种肿瘤组织中增加，包括结肠癌。然而，其在结肠癌中的作用机制仍有待充分阐明。在本研究中，研究了双糖链蛋白聚糖敲低对结肠癌细胞增殖、迁移、侵袭和凋亡的影响。利用RNA干扰技术下调结肠癌细胞系HCT 116中双糖链蛋白聚糖的mRNA表达水平，经G418筛选获得稳定转染的细胞系用于后续实验。结果显示，下调双糖蛋白聚糖的表达抑制细胞增殖，并导致细胞周期停滞在G 0/G1期。Western印迹分析结果还显示，与短发夹RNA对照组相比，沉默biglycan后，细胞周期相关蛋白（包括cyclin A和cyclin D1）的表达水平显著降低，而p21和p27的表达水平显著升高。降低双糖链蛋白聚糖的表达可抑制结肠癌细胞的迁移和侵袭，并诱导细胞凋亡。用SB 203580完全抑制p38信号通路有效地逆转了由双糖链蛋白聚糖下调诱导的凋亡细胞数量的增加。综上所述，本研究的结果表明，双糖蛋白聚糖在结肠癌细胞的增殖、迁移、侵袭和凋亡中发挥重要作用，并且双糖蛋白聚糖通过发挥抗凋亡作用来调节p38 MAPK信号通路。因此，双糖链蛋白聚糖可能是结肠癌基因治疗的一个假定靶点。
Biglycan is an important component of the extracellular matrix, and it is also a member of small leucine-rich proteoglycan family. Previous studies indicated that the expression of biglycan was increased in a variety of tumor tissues, including colon cancer. However, the mechanisms underlying its effects in colon cancer remain to be fully elucidated. In the present study, the effects of biglycan knockdown on colon cancer cell proliferation, migration, invasion and apoptosis were investigated. The mRNA expression levels of biglycan in the HCT116 colon cancer cell line were downregulated using RNA interference, and the stably transfected cell line was obtained through G418 screening for subsequent experiments. The results revealed that downregulation of the expression of biglycan suppressed cell proliferation and caused a cell cycle arrest at the G0/G1 phase. The results of the western blot analysis also revealed that the expression levels of cell cycle-associated proteins, including cyclin A and cyclin D1, were markedly decreased following silencing of biglycan, whereas the expression levels of p21 and p27 were markedly increased compared with that of the-short hairpin RNA control group. Furthermore, the decreased expression of biglycan inhibited colon cancer cell migration and invasion, and induced apoptosis. A complete inhibition of the p38 signaling pathway with SB203580 effectively reversed the increase in apoptotic cell numbers induced by biglycan downregulation. Taken together, the results of the present study indicated that biglycan exerts an important role in cell proliferation, migration, invasion and apoptosis in colon cancer, and that biglycan regulates the p38 MAPK signaling pathway by exerting an antiapoptotic effect. Therefore, biglycan may represent a putative target for colon cancer gene therapy.