Objectives

LncRNAs (long noncoding RNAs) have been reported to critically regulate colorectal cancer (CRC). We prospectively investigated effects and mechanisms of lncRNA LINC00858 on regulation of CRC progression.

Methods

Expression of LINC00858 and its target were analyzed by quantitative real-time polymerase chain reaction and in situ hybridization. MTT and bromodeoxyuridine/5-bromo-2′-deoxyuridine (BrdU) staining to assess cell proliferation ability. Flow cytometry, wound healing, and transwell assays were conducted to evaluate cell apoptosis, migration, and invasion, respectively. Interaction between LINC00858 and its target was confirmed by luciferase activity assay and RNA immunoprecipitation. Subcutaneous xenotransplanted tumor model was established and employed to detect tumorigenic functions of LINC00858, and further evaluated by qRT-PCR, western blot, immunohistochemistry, and hematoxylin and eosin staining.

Results

With a predicted poor prognosis, LINC00858 was upregulated in CRC patients. LINC00858 knockdown suppressed cell proliferation, invasion, and migration abilities, meanwhile induced cell apoptosis. Moreover, LINC00858 could target and inhibit the miR-4766-5p expression, thus promoting CRC progression. miR-4766-5p further suppressed serine/threonine kinase PAK2. Interestingly, interference of LINC00858 suppressed tumorigenic ability of CRC in vivo by downregulating PAK2.

Conclusions

LINC00858 promoted CRC progression by sponging miR-4766 to upregulate PAK2, shedding lights on LINC00858 as a potential therapeutic target candidate in CRC treatment from bench to clinic.

中文翻译：

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LINC00858通过海绵化miR-4766-5p来调节PAK2来促进结直肠癌。

目标

据报道，LncRNA（长非编码RNA）可严格调节结直肠癌（CRC）。我们前瞻性地研究了lncRNA LINC00858对CRC进展调控的作用及其机制。

方法

通过定量实时聚合酶链反应和原位杂交分析了LINC00858及其靶标的表达。MTT和溴脱氧尿苷/ 5-溴-2'-脱氧尿苷（BrdU）染色以评估细胞增殖能力。进行流式细胞术，伤口愈合和transwell分析以分别评估细胞凋亡，迁移和侵袭。荧光素酶活性测定和RNA免疫沉淀证实了LINC00858与它的靶标之间的相互作用。建立皮下异种移植肿瘤模型，并用于检测LINC00858的致瘤功能，并通过qRT-PCR，western印迹，免疫组织化学以及苏木精和曙红染色进一步评估。

结果

预计预后不良，因此CRC患者中LINC00858上调。LINC00858抑制基因抑制细胞增殖，侵袭和迁移能力，同时诱导细胞凋亡。此外，LINC00858可以靶向并抑制miR-4766-5p表达，从而促进CRC进展。miR-4766-5p进一步抑制了丝氨酸/苏氨酸激酶PAK2。有趣的是，LINC00858的干扰通过下调PAK2抑制了CRC在体内的致瘤能力。

结论

LINC00858通过使miR-4766海绵化以上调PAK2来促进CRC的进展，并在LINC00858上点亮了灯，作为从长凳到诊所进行CRC治疗的潜在治疗靶标。