Journal: BMC Cancer

Article Title: METTL3-induced circ_0008345 contributes to the progression of colorectal cancer via the microRNA-182-5p/CYP1A2 pathway

doi: 10.1186/s12885-024-12474-5

Figure Lengend Snippet: circ_0008345 is overexpressed in CRC tissues and cell lines. A PTK2-derived circRNAs were predicted from the circbank database. B RNase R treatment of total RNA and analysis of circ_0008345 and PTK2 mRNA resistance levels. C The expression of circ_0008345 in CRC and adjacent tissues using RT-qPCR. D The circ_0008345 expression in FHC, DLD1, and SW480 cells using RT-qPCR. E Subcellular distribution of circ_0008345. Values are the average of three independent experiments. Data are expressed as the mean ± SD. *** p < 0.001, **** p < 0.0001. p values were determined by paired t-test ( C ), one-way ( D ), or two-way ANOVA ( B )

Article Snippet: Lentiviral vectors harboring short hairpin RNA (shRNA) against circ_0008345 (sh-circ_0008345), METTL3 (sh-METTL3), CYP1A2 (sh-CYP1A2) and their scramble control (sh-NC), miR-182-5p inhibitor and negative control (inhibitor-NC), miR-182-5p mimic and its control (miR-control), circ_0008345 overexpression vector (OE-circ_0008345) and control vector (OE-NC) were all designed and purchased from VectorBuilder (Guangzhou, Guangdong, China).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR

Journal: BMC Cancer

Article Title: METTL3-induced circ_0008345 contributes to the progression of colorectal cancer via the microRNA-182-5p/CYP1A2 pathway

doi: 10.1186/s12885-024-12474-5

Figure Lengend Snippet: circ_0008345 can sponge miR-182-5p. A Search for miRNAs bound to circ_0008345 in circular RNA Interactome. B The expression of circ_0008345 in CRC cells treated with three shRNAs targeting circ_0008345 using RT-qPCR ( n = 3). C Effect of knockdown of circ_0008345 on miR-151-3p and miR-182-5p expression detected by RT-qPCR ( n = 6). D The binding relation between biotinylated miR-151-3p or miR-182-5p and circ_0008345 using a pull-down assay ( n = 3). E Two binding sites of circ_0008345 to miR-182-5p in circular RNA Interactome. F The binding of miR-182-5p and circ_0008345 using a luciferase reporter assay ( n = 3). Data are expressed as the mean ± SD. ** p < 0.01, *** p < 0.001, **** p < 0.0001. p values were determined by two-way ANOVA

Article Snippet: Lentiviral vectors harboring short hairpin RNA (shRNA) against circ_0008345 (sh-circ_0008345), METTL3 (sh-METTL3), CYP1A2 (sh-CYP1A2) and their scramble control (sh-NC), miR-182-5p inhibitor and negative control (inhibitor-NC), miR-182-5p mimic and its control (miR-control), circ_0008345 overexpression vector (OE-circ_0008345) and control vector (OE-NC) were all designed and purchased from VectorBuilder (Guangzhou, Guangdong, China).

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Pull Down Assay, Luciferase, Reporter Assay

Journal: BMC Cancer

Article Title: METTL3-induced circ_0008345 contributes to the progression of colorectal cancer via the microRNA-182-5p/CYP1A2 pathway

doi: 10.1186/s12885-024-12474-5

Figure Lengend Snippet: Inhibition of miR-182-5p rescues the impacts of circ_0008345 knockdown on CRC cell malignant behaviors in vitro and in vivo. CRC cells were treated with sh-circ_0008345 alone or in combination with miR-182-5p inhibitor. A The mRNA and protein expression of CYP1A2 in response to different infections by RT-qPCR and immunoblotting (images of adequate length are shown in the Supplementary File). B The colony formation of CRC cells. C The mobility of CRC cells using Transwell assays. D The viability of CRC cells using EdU assays. E Measurement of tumor volume and weight in mice after subcutaneous injection of transfected DLD1 cells. Values are the average of three independent experiments. Data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. p values were determined by one-way (E) or two-way ANOVA (A-E)

Article Snippet: Lentiviral vectors harboring short hairpin RNA (shRNA) against circ_0008345 (sh-circ_0008345), METTL3 (sh-METTL3), CYP1A2 (sh-CYP1A2) and their scramble control (sh-NC), miR-182-5p inhibitor and negative control (inhibitor-NC), miR-182-5p mimic and its control (miR-control), circ_0008345 overexpression vector (OE-circ_0008345) and control vector (OE-NC) were all designed and purchased from VectorBuilder (Guangzhou, Guangdong, China).

Techniques: Inhibition, In Vitro, In Vivo, Expressing, Quantitative RT-PCR, Western Blot, Injection, Transfection

Journal: BMC Cancer

Article Title: METTL3-induced circ_0008345 contributes to the progression of colorectal cancer via the microRNA-182-5p/CYP1A2 pathway

doi: 10.1186/s12885-024-12474-5

Figure Lengend Snippet: miR-182-5p alters CRC cell growth and motility by binding to CYP1A2. A The mRNA expression of CYP1A2 in response to different infections by RT-qPCR. CRC cells were treated with OE-circ_0008345 alone or in combination with sh-CYP1A2. B The colony formation of CRC cells. C The migration and invasion of CRC cells using Transwell assays. D The viability of CRC cells using EdU assays. E Measurement of tumor volume and weight in mice after subcutaneous injection of transfected DLD1 cells. CRC cells were treated with miR-182-5p inhibitor alone or in combination with sh-CYP1A2. F The colony formation of CRC cells. G The migration and invasion of CRC cells using Transwell assays. H The viability of CRC cells using EdU assays. I Measurement of tumor volume and weight in mice after subcutaneous injection of transfected DLD1 cells. Values are the average of three independent experiments or six mice. Data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. p values were determined by one-way or two-way ANOVA

Article Snippet: Lentiviral vectors harboring short hairpin RNA (shRNA) against circ_0008345 (sh-circ_0008345), METTL3 (sh-METTL3), CYP1A2 (sh-CYP1A2) and their scramble control (sh-NC), miR-182-5p inhibitor and negative control (inhibitor-NC), miR-182-5p mimic and its control (miR-control), circ_0008345 overexpression vector (OE-circ_0008345) and control vector (OE-NC) were all designed and purchased from VectorBuilder (Guangzhou, Guangdong, China).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Migration, Injection, Transfection

Journal: BMC Cancer

Article Title: METTL3-induced circ_0008345 contributes to the progression of colorectal cancer via the microRNA-182-5p/CYP1A2 pathway

doi: 10.1186/s12885-024-12474-5

Figure Lengend Snippet: METTL3 mediates circ_0008345 upregulation in CRC cells. A The presence of m6A modification site in circ_0008345 predicting using circbank. B Detection of m6A-modified RNA levels in cells by MeRIP. C The effect of three shRNAs knocking down METTL3 using RT-qPCR. D Detection of circ_0008345 expression after knockdown of METTL3 using RT-qPCR. E Stability level of circ_0008345 in actinomycin D-treated DLD1 and SW480 cells. Values are the average of three independent experiments. Data are expressed as the mean ± SD. **** p < 0.0001. p values were determined by one-way or two-way ANOVA

Article Snippet: Lentiviral vectors harboring short hairpin RNA (shRNA) against circ_0008345 (sh-circ_0008345), METTL3 (sh-METTL3), CYP1A2 (sh-CYP1A2) and their scramble control (sh-NC), miR-182-5p inhibitor and negative control (inhibitor-NC), miR-182-5p mimic and its control (miR-control), circ_0008345 overexpression vector (OE-circ_0008345) and control vector (OE-NC) were all designed and purchased from VectorBuilder (Guangzhou, Guangdong, China).

Techniques: Modification, Quantitative RT-PCR, Expressing

Journal: BMC Cancer

Article Title: METTL3-induced circ_0008345 contributes to the progression of colorectal cancer via the microRNA-182-5p/CYP1A2 pathway

doi: 10.1186/s12885-024-12474-5

Figure Lengend Snippet: Schematic illustration of circ_0008345 expediting the progression of CRC. METTL3-mediated m6A modification in CRC can upregulate circ_0008345. circ_0008345, as a CYP1A2 ceRNA, can act as a “sponge” and interact with miR-182-5p to increase the mRNA expression of CYP1A2

Article Snippet: Lentiviral vectors harboring short hairpin RNA (shRNA) against circ_0008345 (sh-circ_0008345), METTL3 (sh-METTL3), CYP1A2 (sh-CYP1A2) and their scramble control (sh-NC), miR-182-5p inhibitor and negative control (inhibitor-NC), miR-182-5p mimic and its control (miR-control), circ_0008345 overexpression vector (OE-circ_0008345) and control vector (OE-NC) were all designed and purchased from VectorBuilder (Guangzhou, Guangdong, China).

Techniques: Modification, Expressing