Journal: Cancer Science

Article Title: Circular RNA circ0104103 inhibits colorectal cancer progression through interactions with HuR and miR ‐373‐5p

doi: 10.1111/cas.15695

Figure Lengend Snippet: Expression and characterization of Circ0104103 in colorectal cancer (CRC). (A) Schematic illustration demonstrating the structure of Circ_0104013 and Alu elements in the flanking sequence. (B) RT‐PCR identified the presence of Circ0104103 in the HCT116 cells. Divergent primers amplified Circ0104103 in cDNA but not in genomic DNA. GAPDH was used as the negative control. (C, D) Quantitative RT‐PCR (qRT‐PCR) detected the expression of Circ0104103 and lin0104103 in CRC cells with or without RNase R or actinomycin D treatment. (E) Relative Circ0104103 expression levels in nuclear and cytosolic fractions of HCT116 cells. Nuclear controls: U6, cytosolic controls: GAPDH. (F) FISH assay identifying the subcellular location of Circ0104103 in the HCT116 cells. (G, H) qRT‐PCR for the expression of Circ0104103 in CRC cell lines and tissues. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human normal colorectal epithelial cell line NCM460 and the human CRC cell lines (HCT116, HT29, HCT8, SW480, SW620, DLD1 and CACO2) were obtained from the ATCC.

Techniques: Expressing, Sequencing, Reverse Transcription Polymerase Chain Reaction, Amplification, Negative Control, Quantitative RT-PCR

Journal: Cancer Science

Article Title: Circular RNA circ0104103 inhibits colorectal cancer progression through interactions with HuR and miR ‐373‐5p

doi: 10.1111/cas.15695

Figure Lengend Snippet: Circ0104103 inhibits colorectal cancer (CRC) cell growth and metastasis in vitro. (A) HCT116 and HCT8 cells infected with circ0104103 or sh_circ0104103#2 were seeded into six‐well plates. The number of colonies was counted on the 14th day after seeding. (B) EdU assays were used to determine the cell proliferation ability of infected cells. (C) Representative images of wound healing assays performed using HCT116 cells and HCT8 cells after Circ0104103 was silenced or overexpressed. (D)Transwell assays were used to determine the invasion abilities of infected cells. (E) Immunoblot analysis of MMP9, PCNA, BCL2, and P53 protein expression in circ0104103‐overexpressing HCT116 cells and circ0104103 silencing HCT116 cells. Scale bar = 100 μm. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human normal colorectal epithelial cell line NCM460 and the human CRC cell lines (HCT116, HT29, HCT8, SW480, SW620, DLD1 and CACO2) were obtained from the ATCC.

Techniques: In Vitro, Infection, Western Blot, Expressing

Journal: Cancer Science

Article Title: Circular RNA circ0104103 inhibits colorectal cancer progression through interactions with HuR and miR ‐373‐5p

doi: 10.1111/cas.15695

Figure Lengend Snippet: Circ0104103 inhibits colorectal cancer (CRC) cell growth in vivo. (A) Representative image of tumors formed in nude mice from empty vector, sh‐ circ0104103#2 vector, and circ0104103 overexpression vector groups and the tumor volume growth curves and weight of different groups. (B) Representative H&E staining and the number of lung metastatic nodules in nude mice in the three groups. (C) Representative images for H&E‐staining, BCL2, MMP9, P53, and PCNA immunostaining of tumor samples from the different groups. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human normal colorectal epithelial cell line NCM460 and the human CRC cell lines (HCT116, HT29, HCT8, SW480, SW620, DLD1 and CACO2) were obtained from the ATCC.

Techniques: In Vivo, Plasmid Preparation, Over Expression, Staining, Immunostaining

Journal: Cancer Science

Article Title: Circular RNA circ0104103 inhibits colorectal cancer progression through interactions with HuR and miR ‐373‐5p

doi: 10.1111/cas.15695

Figure Lengend Snippet: Circ0104103 enhances beta‐lactamase‐like (LACTB) expression in colorectal cancer (CRC). (A, B) Quantitative RT‐PCR (qRT‐PCR) and western blot assays detected the expression of LACTB in HCT116 and HCT8 cell lines with circ0104103 knockdown or overexpression. (C) LACTB expression was detected by immunohistochemistry (IHC) xenograft tumor tissues. LACTB protein levels in CRC tissues and corresponding normal tissues was detected by IHC. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human normal colorectal epithelial cell line NCM460 and the human CRC cell lines (HCT116, HT29, HCT8, SW480, SW620, DLD1 and CACO2) were obtained from the ATCC.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Over Expression, Immunohistochemistry, Paraffin-embedded Immunohistochemistry

Journal: Cancer Science

Article Title: Circular RNA circ0104103 inhibits colorectal cancer progression through interactions with HuR and miR ‐373‐5p

doi: 10.1111/cas.15695

Figure Lengend Snippet: Circ0104103 upregulates beta‐lactamase‐like (LACTB) expression by sponging miR‐373‐5p. (A) Colony formation assays of Circ0104103 overexpressed colorectal cancer (CRC) cells with transfection of miR‐373‐5p mimics. (B) Representative images of EdU assays performed using HCT116 cells with indicated treatment. (C) Representative images of Transwell invasion assays performed using CRC cells with indicated treatment. (D) Representative images of wound healing assays performed using HCT116 cells with indicated treatment. Scale bar = 100 μm. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human normal colorectal epithelial cell line NCM460 and the human CRC cell lines (HCT116, HT29, HCT8, SW480, SW620, DLD1 and CACO2) were obtained from the ATCC.

Techniques: Expressing, Transfection

Journal: Cancer Science

Article Title: Circular RNA circ0104103 inhibits colorectal cancer progression through interactions with HuR and miR ‐373‐5p

doi: 10.1111/cas.15695

Figure Lengend Snippet: Tumor‐suppressing functions of Circ0104103 are dependent on beta‐lactamase‐like (LACTB). (A) Colony formation assays demonstrated that overexpression of circ0104103 inhibited cancer cell growth. LACTB knockdown could abolish growth inhibition caused by circ0104103. (B) EdU assays showed that LACTB knockdown abolished the decreased proliferation rates of colorectal cancer (CRC) cells caused by circ0104103. (C, D) Transwell assays and wound healing assays demonstrated that LACTB knockdown abolished the decreased abilities of migration and invasion caused by circ0104103. (E) Immunoblot analysis of MMP9, PCNA, BCL2, and P53 protein expression in HCT116 cells with indicated treatment. (F) Schematic of the proposed mechanism of circ0104103 in CRC. Scale bar = 100 μm. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human normal colorectal epithelial cell line NCM460 and the human CRC cell lines (HCT116, HT29, HCT8, SW480, SW620, DLD1 and CACO2) were obtained from the ATCC.

Techniques: Over Expression, Knockdown, Inhibition, Migration, Western Blot, Expressing