NFE2L3 Inhibition Induces Cell Cycle Arrest at the G0/G1 Phase in Colorectal Cancer Cells through Downregulating CCND1 and pRb1-ser807/811

Abstract

The molecular mechanism for colorectal cancer to develop remains unelucidated. To find biomarkers related to colorectal cancer development, we analyzed the gene expression profile of 380 colorectal cancer patients and 51 healthy controls by R software. Finally, 1579 upregulated differential expression genes (DEGs) and 3218 downregulated DEGs were identified. Then, the top 20 upregulated DEGs were compared with 181 upregulated DEGs that we reported previously, and 11 overlapped DEGs were found. NFE2L3 (nuclear factor, erythroid 2-like 3) was among those overlapped DEGs and was rarely reported in colorectal cancer. Real-time polymerase chain reaction (PCR) results showed that higher NFE2L3 expression levels were identified in paired tumor samples than in paratumor samples (48 paired samples). Flow cytometry analysis revealed that the cell cycle was arrested at the G0/G1 phase after inhibition of NFE2L3 in both HCT116 and SW480 cell lines. Western blot detection showed that CCND1 and phosphorylated Rb transcriptional corepressor 1 at ser-807/811 (pRb1-ser807/811) expression levels were downregulated when NFE2L3 was inhibited in those two cell lines. A significant positive correlation was observed between NFE2L3 and CCND1 expression levels in colorectal tissue samples. These evidences indicate that downregulation of NFE2L3 induces cell cycle arrest at the G0/G1 phase through downregulation of CCND1 and pRb1-ser807/811.

NFE2L3 promotes malignant behavior and EMT of human hepatocellular carcinoma (HepG2) cells via Wnt/β‑catenin pathway

“…Several studies have reported that decreased NFE2L3 expression inhibits proliferation of cells 15 , 18 . To investigate the potential effect of NFE2L3 on HepG2 cell proliferation, NFE2L3 shRNA interference lentiviral vector was performed to knockdown NFE2L3.…”

“…Consistent with the above results, the expression of CDK2 and PCNA also decreased, it is further confirmed that NFE2L3 plays an important role in cell growth and proliferation. Cyclin D1 is a key mediator of cell growth and proliferation, some researches had proved that NFE2L3 regulated the expression of cyclin D1 in CRC while another article pointed out that the silencing of NFE2L3 will increase the level of DUX4, a direct inhibitor of CDK1, and reduce the proliferation and tumorigenicity of colon cancer cells 15 , 18 . In addition, both cytoplasmic and nuclear β-catenin were decreased by knockdown NFE2L3, indicating NFE2L3 significantly regulated transcription of β-catenin in liver cancer cells.…”

“…The survival rate of patients with colorectal cancer is extremely low in those with high expression of NFE2L3. Zhang et al showed that the absence of NFE2L3 induces cell cycle arrest at the G0/G1 phase in colorectal cancer cells 15 . These findings suggest that NFE2L3 may be a key regulator of cancer progression.…”

SNRPD1 confers diagnostic and therapeutic values on breast cancers through cell cycle regulation

“…8A). We, in addition, tested the expression of CCND1 whose down-regulation is associated with G 0 /G 1 arrest [28] but missed from the dataset we used for SRFG identi cation. CCND1 was down-regulated to approximately 20% and 60% of the control in SNPRD1-silenced MCF7 and MDAMB231 cells at both gene and protein expression levels (p = 0.022 at the gene expression level, p = 1.53E-4 at the protein expression level in MCF7, p = 0.0023 at the gene expression level, p = 1.7E-166 at the protein expression level in MDAMB231, Fig.…”

“…Downward-regulated expression of CDCA5 [29,30], NDC80 [31], CCNA2 [32] on SNRPD1 silencing suggested reduced synthesis of DNA and proteins as well as recessed cell mitosis. Further evidence on reduced CCND1 expression after knocking down SNRPD1 implicated the accumulation of cells in the G 0 /G 1 state given the regulatory role of CCND1 in triggering G /G 1 cell cycle arrest [28].…”