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Effect of long non-coding RNA Gas5 on proliferation, migration, invasion and apoptosis of colorectal cancer HT-29 cell line

机译:长非编码RNA Gas5对结直肠癌HT-29细胞增殖,迁移,侵袭和凋亡的影响

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This study aims to investigate the effect of long non-coding RNA (lncRNA) Gas5 on proliferation, migration, invasion and apoptosis of colorectal cancer (CRC) HT-29 cell line. CRC and normal tissues were collected and prepared from a total of 126 CRC patients, and normal intestinal epithelial cell line FHC and CRC cell lines (HCT-8, HT-29, HCT-116 and SW-480) were prepared. Gas5 expression was detected by quantitative reverse transcriptase-polymerase chain reaction. HT-29 cell line exhibiting the lowest Gas5 expression was selected for further experimentation and divided into blank, negative control and pcNDA-Gas5 groups. The cell counting kit-8 assay was used to test cell proliferation. Flow cytometry was applied to examine cell apoptosis. Transwell assay was performed to detect the migration and invasion of HT-29 cells. The mRNA and protein expression of factors in the classical proliferation (Akt/Erk) and apoptosis (caspase-9/caspase-3) pathways were detected. Gas5 expression was lower in CRC tissues compared to the adjacent normal tissues, and is also lower in CRC cell lines than FHC cell line. Gas5 expression was associated with tumor size and TNM staging. Gas5 expression, distant metastasis, tumor differentiation and TNM staging were independent CRC prognostic factors. The results showed that elevated Gas5 expression inhibited proliferation, migration and invasion, but promoted apoptosis of CRC cells. Meanwhile, elevated Gas5 expression inhibited mRNA expression of Akt and Erk and protein expression of p-Akt and p-Erk, which promoted Casp9 mRNA and pho-Casp9 protein expression but inhibited Casp3 mRNA and pho-Casp3 protein expression. The findings indicated that overexpression of lncRNA Gas5 can inhibit the proliferation, migration and invasion but promote apoptosis of CRC cells.
机译:这项研究旨在研究长非编码RNA(lncRNA)Gas5对结直肠癌(CRC)HT-29细胞系增殖,迁移,侵袭和凋亡的影响。从总共126名CRC患者中收集并制备CRC和正常组织,并制备了正常肠上皮细胞系FHC和CRC细胞系(HCT-8,HT-29,HCT-116和SW-480)。通过定量逆转录酶-聚合酶链反应检测Gas5表达。选择表现出最低的Gas5表达的HT-29细胞系进行进一步实验,并分为空白,阴性对照和pcNDA-Gas5组。细胞计数试剂盒8测定法用于测试细胞增殖。流式细胞仪用于检查细胞凋亡。进行Transwell测定以检测HT-29细胞的迁移和侵袭。检测到经典增殖(Akt / Erk)和凋亡(caspase-9 / caspase-3)途径中因子的mRNA和蛋白表达。与相邻的正常组织相比,CRC组织中的Gas5表达更低,并且在CRC细胞系中也比FHC细胞系更低。 Gas5表达与肿瘤大小和TNM分期有关。 Gas5表达,远处转移,肿瘤分化和TNM分期是独立的CRC预后因素。结果表明,高表达的Gas5抑制增殖,迁移和侵袭,但促进CRC细胞凋亡。同时,Gas5表达升高抑制Akt和Erk的mRNA表达以及p-Akt和p-Erk的蛋白表达,从而促进Casp9 mRNA和pho-Casp9蛋白的表达,但抑制Casp3 mRNA和pho-Casp3蛋白的表达。这些发现表明,lncRNA Gas5的过表达可以抑制CRC细胞的增殖,迁移和侵袭,但促进其凋亡。

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