The mechanism of m6Am-modifying enzyme PCIF1 regulating target gene ACOT8 in gastric cancer progression

doi:10.19401/j.cnki.1007-3639.2023.04.007

Abstract

Abstract:

Background and purpose: Recent evidence suggests that N6, 2'-O- dimethyladenosine (m6Am) modifying enzyme phosphorylated C-terminal domain-interacting factor 1 (PCIF1) may be an important biomarker/therapeutic target in gastric cancer. However, the relationship between this novel PCIF1 molecular mechanism and gastric cancer progression remains to be further explored. In this study, we analyzed the regulatory role of PCIF1 on gastric cancer proliferation, migration and invasion and the mechanism of its regulatory target gene ACOT8 in gastric cancer progression. Methods: PCIF1 expression in gastric cancer and non-gastric cancer tissues of gastric cancer patients was analyzed using Gene Expression Profile Interaction Analysis (GEPIA), and the correlation of PCIF1 expression with overall survival in gastric cancer patients was analyzed. We collected all gastric cancer tissues and matched para-cancerous tissues from 89 primary gastric cancer patients treated in Department of Gastroenterology of Changsha First Hospital from 2019 to 2021. PCIF1 expression was analyzed by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. In in vitro experiments, SNU5 cells were divided into PCIF1 knockdown (sh-PCIF1) group and corresponding control (sh-NC) group, and AGS cells were divided into vehicle as normal control (NC) group and PCIF1 overexpression (PCIF1) group. The effect of PCIF1 on gastric cancer cell proliferation, invasion and migration was investigated using cell counting kit-8 (CCK-8), EdU and transwell assays. Furthermore, ACOT8 expression was knocked down in PCIF1-overexpressing AGS cells, and rescue experiments were performed. A subcutaneous xenograft model was used to determine the biological effects of PCIF1 in gastric cancer in vivo. Results: PCIF1 expression was abnormally high in gastric cancer tissues and cell lines, with significant correlation between PCIF1 expression and the prognosis of gastric cancer patients. Compared with the sh-NC group, the cell viability, EdU positive cells, migration and invasion cell numbers in the sh-PCIF1 group were significantly decreased (P<0.05). Compared with the NC group, the cell viability, EdU positive cells, migration and invasion cell numbers in the PCIF1 group were significantly increased (P<0.05). In PCIF1-overexpressing AGS cells, knockdown of ACOT8 expression decreased cell viability, EdU-positive cells and migrating and invasive cell numbers. In vivo experiments showed, compared with the NC group, the tumor volume and weight of the nude mice in the PCIF1-overexpressing group were significantly increased (P<0.05). Conclusions: PCIF1 is upregulated in gastric cancer cell lines and tissues. In addition, the PCIF1/ACOT8 axis is involved in mediating the malignant behavior of gastric cancer cells.

Key words: N6, 2′-O-dimethyladenosine, Phosphorylated C-terminal domain-interacting factor 1, Acyl-CoA thioesterase 8, Gastric cancer, Proliferation, Metastasis

CLC Number:

R735.2

PENG Jin, WANG Weining, TAN Zhi, YE Guannan, ZHOU Zhen. The mechanism of m6Am-modifying enzyme PCIF1 regulating target gene ACOT8 in gastric cancer progression[J]. China Oncology, 2023, 33(4): 368-376.

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