长链非编码RNA(lncRNA)的失调,如母本表达基因3(MEG3)和长基因间非编码RNA重编程调节因子(linc-ROR),在结直肠癌的进展中起着至关重要的作用。我们旨在同时评估linc-ROR沉默和MEG3激活对结直肠癌细胞增殖的影响;并探索TP53相关通路的潜在机制。在双向CEA启动子(UM1)下克隆MEG3和linc-RORshRNA。随后,构建另外的载体以表达linc-RORshRNA(UM2)和MEG3(UM3)。用这些重组载体转染结直肠癌细胞系后,细胞活力实验,凋亡,进行细胞周期分析。此外,使用定量实时聚合酶链反应(qRT-PCR)评估TP53的转录活性和相关基因。有趣的是,UM1比UM2和UM3显著抑制两种细胞系的增殖。响应于UM1,在HCT116细胞中TP53转录物(10.46)比SW480细胞(6.16)显着增加;这导致TP53INP1,TP53I3,GDF15,CCKN1A和BAX的上调,和G1细胞周期蛋白(D1,E1)的下调。HCT116(36.35%)和SW480(16.64%)细胞的凋亡率比对照组增加。此外,UM1转染的HCT116细胞在G0/G1期表现出明显的停滞,伴随着G2/M细胞群体的减少。与单向向量相比,同时靶向方法在转录水平上增强了TP53的激活.细胞对UM1的反应导致TP53的快速上调,导致细胞增殖的抑制,细胞凋亡增加,和细胞周期停滞。这些发现表明,靶向MEG3和linc-ROR的协同作用可以作为TP53相关结肠癌的有希望的治疗策略。
Dysregulation of long noncoding RNAs (lncRNAs), such as maternally expressed gene 3 (MEG3) and long intergenic noncoding RNA regulator of reprogramming (linc-ROR), plays a crucial role in colorectal cancer progression. We aimed to assess linc-ROR silencing and MEG3 activation on the colorectal cancer cell proliferation simultaneously; and explore the underlying mechanisms in the
TP53-associated Pathway. The MEG3 and linc-ROR shRNA were cloned under the bidirectional CEA promoter (UM1). Subsequently, additional vectors were constructed to express linc-ROR shRNA (UM2) and MEG3 (UM3). After transfecting colorectal cancer cell lines with these recombinant vectors, experiments on cell viability, apoptosis, and cell cycle analysis were conducted. Furthermore,
TP53\'s transcriptional activity and associated genes were assessed using quantitative real-time polymerase chain reaction (qRT-PCR). Interestingly, UM1 significantly inhibited the proliferation of both cell lines than UM2 and UM3. In response to UM1,
TP53 transcript remarkably increased in HCT116 cells (10.46) than SW480 cells (6.16); which resulted in up-regulation of TP53INP1, TP53I3, GDF15, CCKN1A and BAX, and down-regulation of G1 cyclins (D1, E1). The rate of apoptosis increased in HCT116 (36.35 %) and SW480 (16.64 %) cells than control. Moreover, UM1-transfected HCT116 cells exhibited a notable arrest in the G0/G1 phase, accompanied by a reduction in the G2/M cell population. Compared to unidirectional vectors, the concurrent targeting approach enhanced
TP53 activation at the transcription level. The cell response to UM1 resulted in rapid upregulation of
TP53, leading to inhibition of cell proliferation, increased apoptosis, and cell cycle arrest. These findings suggest that the synergistic effect of targeting both MEG3 and linc-ROR could serve as a promising therapeutic strategy for TP53-associated colon cancer.