长链非编码 RNA 00337 的沉默通过 MEK/ERK 通路抑制结直肠癌细胞的增殖、细胞周期进程、迁移和侵袭。
Silencing of linc00337 inhibits proliferation, cell cycle progression, migration, and invasion of colorectal cancer cells through the MEK/ERK pathway.
机构信息
Department of General Surgery, Jiangdu People's Hospital Affiliated to Medical College of Yangzhou University, Yangzhou, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 May;24(10):5353-5359. doi: 10.26355/eurrev_202005_21318.
OBJECTIVE
To explore the expression and biological functions of linc00337 in colorectal cancer (CRC), as well as its underlying mechanism.
PATIENTS AND METHODS
The relative expression of linc00337 in 47 cases of CRC tissues and cells was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The si-linc00337 interference sequences were designed and transiently transfected into CRC cells. The interference efficiency was detected via qRT-PCR. Regulatory effect of linc00337 on proliferation of CRC cells was detected via colony formation assay. Cell cycle distribution and apoptosis rate after interference in linc00337 expression were determined using flow cytometry. Moreover, the effects of linc00337 knockdown on cell migration and invasion were detected using transwell assay. At last, the effect of si-linc00337 on the MEK/ERK signaling pathway was detected using Western blotting.
RESULTS
The results of qRT-PCR showed that among the 47 cases of CRC tissues, the expression of linc00337 was up-regulated in 40 cases. Similarly, it was highly expressed in CRC cell lines. The results of colony formation assay manifested that cell proliferation declined after interference in linc00337 expression. The results of flow cytometry and transwell assay showed that interference in linc00337 expression arrested the cell cycle in G1/G0 phase, increased the apoptosis rate, and inhibited the invasion and migration of CRC cells. According to the results of Western blotting, expressions of molecular markers in the MEK/ERK pathway after interference in linc00337 expression were significantly changed.
CONCLUSIONS
Linc00337 is up-regulated in CRC tissues and cells. Interference in linc00337 expression can inhibit cell proliferation, migration, and invasion and promote apoptosis through the MEK/ERK pathway.
目的
探讨 linc00337 在结直肠癌（CRC）中的表达及生物学功能及其潜在机制。
方法
采用实时定量逆转录聚合酶链反应（qRT-PCR）检测 47 例 CRC 组织和细胞中 linc00337 的相对表达。设计并瞬时转染 si-linc00337 干扰序列至 CRC 细胞，qRT-PCR 检测干扰效率。采用集落形成实验检测 linc00337 对 CRC 细胞增殖的调控作用。通过流式细胞术检测干扰 linc00337 表达后细胞周期分布和凋亡率。此外，通过 Transwell 实验检测 linc00337 敲低对细胞迁移和侵袭的影响。最后，通过 Western blot 检测 si-linc00337 对 MEK/ERK 信号通路的影响。
结果
qRT-PCR 结果显示，在 47 例 CRC 组织中，40 例组织中 linc00337 表达上调。同样，在 CRC 细胞系中也呈高表达。集落形成实验结果表明，干扰 linc00337 表达后细胞增殖能力下降。流式细胞术和 Transwell 实验结果表明，干扰 linc00337 表达可使细胞周期停滞在 G1/G0 期，增加细胞凋亡率，抑制 CRC 细胞的侵袭和迁移。Western blot 结果显示，干扰 linc00337 表达后 MEK/ERK 通路中分子标志物的表达明显改变。
结论
linc00337 在 CRC 组织和细胞中上调。干扰 linc00337 表达可通过 MEK/ERK 通路抑制细胞增殖、迁移和侵袭，促进细胞凋亡。
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