Department of Medical Ultrasound, Xuzhou Hospital Affiliated to School of Medicine, Southeast University, Xuzhou, China.
Eur Rev Med Pharmacol Sci. 2018 Mar;22(5):1358-1365. doi: 10.26355/eurrev_201803_14479.
We investigated the expression of human long non-coding ribonucleic acid (lncRNA), BRAF-activated non-coding RNA (BANCR) in breast cancer tissues and its effects on the in vitro proliferation, apoptosis, invasion and metastasis of breast cancer cells; also, we investigated its possible mechanism.
The expressions of BANCR in 65 pairs of breast cancer tissues and para-carcinoma normal breast tissues were detected by Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The expressions of BANCR in normal breast epithelial cells (MCF10A) and breast cancer cells (MCF-7, MDA-MB-231, SKBR3 and BT-20) were further detected. The knockdown efficiency of BANCR small interfering RNA (siRNA) in MCF-7 cells was detected by qRT-PCR. The effects of BANCR knockdown on proliferation, apoptosis, invasion and metastasis capacities of MCF-7 cells were explored via methyl thiazolyl tetrazolium (MTT) proliferation assay, cell colony formation assay, fluorescence-activated cell sorting (FACS) and transwell migration assay. Western blotting was used to detect the changes in expressions of apoptosis-related proteins, epithelial-mesenchymal transition (EMT)-related proteins and matrix metalloproteinases (MMPs) after knockdown of BANCR.
The expression level of lncRNA BANCR in breast cancer tissues was significantly higher than that in para-carcinoma normal tissues. The prognosis of patients in low-expression BANCR group was significantly superior to that of patients in high-expression BANCR group. After BANCR knockdown in breast cancer MCF-7 cells, the cell proliferation and colony formation capacities were significantly inhibited. Further mechanism research showed that inhibiting BANCR could promote the apoptosis of MCF-7 cells. Results of Western blotting revealed that the expressions of B-cell lymphoma 2 associated X protein (BAX), cleaved-Caspase-3 and cleaved-poly adenosine diphosphate-ribose polymerase (PARP) in knockdown group were significantly up-regulated compared with those in control group. Both wound-healing assay and transwell migration assay showed that the down-regulation of lncRNA BANCR could inhibit the invasion and metastasis capacities of MCF-7 cells, whose mechanism was related to the inhibition of EMT process and down-regulation of MMP expressions in cells.
LncRNA BANCR is highly expressed in breast cancer, which is significantly correlated with the prognosis of patients; moreover, it can promote the growth, invasion and metastasis of ovarian cancer cells. The down-regulation of BANCR can inhibit the proliferation, invasion and metastasis capacities of MCF-7 cells.
我们研究了人类长链非编码 RNA(lncRNA)、BRAF 激活的非编码 RNA(BANCR)在乳腺癌组织中的表达及其对乳腺癌细胞体外增殖、凋亡、侵袭和转移的影响;还探讨了其可能的机制。
采用实时荧光定量聚合酶链反应(qRT-PCR)检测 65 对乳腺癌组织和癌旁正常乳腺组织中 BANCR 的表达。进一步检测正常乳腺上皮细胞(MCF10A)和乳腺癌细胞(MCF-7、MDA-MB-231、SKBR3 和 BT-20)中 BANCR 的表达。qRT-PCR 检测 BANCR 小干扰 RNA(siRNA)在 MCF-7 细胞中的敲低效率。通过甲基噻唑基四唑(MTT)增殖试验、细胞集落形成试验、荧光激活细胞分选(FACS)和 Transwell 迁移试验探讨 BANCR 敲低对 MCF-7 细胞增殖、凋亡、侵袭和转移能力的影响。Western blot 检测 BANCR 敲低后凋亡相关蛋白、上皮-间充质转化(EMT)相关蛋白和基质金属蛋白酶(MMPs)表达的变化。
乳腺癌组织中 lncRNA BANCR 的表达水平明显高于癌旁正常组织。BANCR 低表达组患者的预后明显优于 BANCR 高表达组患者。乳腺癌 MCF-7 细胞中 BANCR 敲低后,细胞增殖和集落形成能力明显受到抑制。进一步的机制研究表明,抑制 BANCR 可促进 MCF-7 细胞凋亡。Western blot 结果显示,敲低组 B 细胞淋巴瘤 2 相关 X 蛋白(BAX)、cleaved-Caspase-3 和 cleaved-poly 腺苷二磷酸核糖聚合酶(PARP)的表达明显高于对照组。划痕愈合试验和 Transwell 迁移试验均表明,下调 lncRNA BANCR 可抑制 MCF-7 细胞的侵袭和转移能力,其机制与 EMT 过程的抑制和细胞中 MMP 表达的下调有关。
lncRNA BANCR 在乳腺癌中高表达,与患者预后显著相关;此外,它可以促进卵巢癌细胞的生长、侵袭和转移。BANCR 的下调可抑制 MCF-7 细胞的增殖、侵袭和转移能力。