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长链非编码 RNA BANCR 表明乳腺癌预后不良,并促进细胞增殖和侵袭。

Long non-coding RNA BANCR indicates poor prognosis for breast cancer and promotes cell proliferation and invasion.

机构信息

Department of Medical Ultrasound, Xuzhou Hospital Affiliated to School of Medicine, Southeast University, Xuzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2018 Mar;22(5):1358-1365. doi: 10.26355/eurrev_201803_14479.

Abstract

OBJECTIVE

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.

PATIENTS AND METHODS

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.

RESULTS

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.

CONCLUSIONS

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.

摘要

目的

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。

患者和方法

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。

结果

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。

结论

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。

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