Journal: Pharmacogenomics and Personalized Medicine

Article Title: Hsa_circ_0078767 Enhances Osteosarcoma Chemoresistance to Doxorubicin Through the Regulation of the miR-188-3p/GPX4 Axis

doi: 10.2147/PGPM.S473702

Figure Lengend Snippet: DOX-resistant OS tumors and cell lines express higher levels of hsa_circ_0078767. ( A ) qPCR was used to detect hsa_circ_0078767 levels in 65 and 61 non-respond and respond OS tissue samples, respectively. ( B ) Kaplan-Meier curves for assessment of the association between OS patient survival andhsa_circ_0078767 levels. ( C ) hsa_circ_0078767 expression in parental (HOS, U2OS) and DOX-resistant (HOS/DOX, U2OS/DOX) OS cell lines, shown by qPCR. ( D and E ) RNase R was used to treat U2OS/DOX and HOS/DOX cells, after which qPCR was utilized to assess hsa_circ_0078767 and DYNC1H1 expression. ( F ) Hsa_circ_0078767, GAPDH, and U6 were detected from the nuclear and cytoplasmic fractions of U2OS/DOX and HOS/DOX cells. ( G ) hsa_circ_0078767 and linear DYNC1H1 transcript stability levels were analyzed in U2OS/DOX and HOS/DOX cells. *P < 0.05.

Article Snippet: The KHOS and U2OS human OS cell lines from Biovector (Beijing, China) were grown in DMEM (Gibco, MA, USA) supplemented with 10% FBS (Gibco) and penicillin/streptomycin (Invitrogen, MA, USA) in a 5% CO 2 37°C incubator.

Techniques: Expressing

Journal: Pharmacogenomics and Personalized Medicine

Article Title: Hsa_circ_0078767 Enhances Osteosarcoma Chemoresistance to Doxorubicin Through the Regulation of the miR-188-3p/GPX4 Axis

doi: 10.2147/PGPM.S473702

Figure Lengend Snippet: Knocking down hsa_circ_0078767 reduces in vitro OS cell resistance to DOX. ( A and B ) qPCR was employed to gauge knockdown efficiency in U2OS/DOX and HOS/DOX cells following si-NC or si-hsa_circ_0078767 transfection. ( C ) CCK-8 assays were utilized to calculate IC 50 values for DOX-treated OS cells. *P < 0.05.

Article Snippet: The KHOS and U2OS human OS cell lines from Biovector (Beijing, China) were grown in DMEM (Gibco, MA, USA) supplemented with 10% FBS (Gibco) and penicillin/streptomycin (Invitrogen, MA, USA) in a 5% CO 2 37°C incubator.

Techniques: In Vitro, Knockdown, Transfection, CCK-8 Assay

Journal: Pharmacogenomics and Personalized Medicine

Article Title: Hsa_circ_0078767 Enhances Osteosarcoma Chemoresistance to Doxorubicin Through the Regulation of the miR-188-3p/GPX4 Axis

doi: 10.2147/PGPM.S473702

Figure Lengend Snippet: miR-188-3p is targeted directly by hsa_circ_0078767. ( A ) Putative hsa_circ_0078767 and miR-188-3p binding sites. ( B ) miR-188-3p levels in 65 and 61 non-respond and respond OS tissue samples, respectively, shown by qPCR. ( C ) Spearman correlation coefficients between hsa_circ_0078767 and miR-188-3p levels in DOX-resistant OS patient tissue samples. ( D and E ) Dual-luciferase reporter assays for assessment of HOS/DOX and U2OS/DOX cells following hsa_circ_0078767-wt or hsa_circ_0078767-mut co-transfection with the miR-188-3p or miR-NC constructs. ( F and G ) Interaction betweenhsa_circ_0078767 and miR-188-3p shown by RIP. ( H ) Interaction between hsa_circ_0078767 and miR-188-3p shown by RNA pull-down. ( I ) miR-188-3p levels in parental and DOX-resistant OS cells. ( J and K ) qPCR determination of miR-188-3p expression in U2OS/DOX and HOS/DOX cells following si-NC, si-hsa_circ_0078767, pCD-ciR, or hsa_circ_0078767 transfection. *P < 0.05.

Article Snippet: The KHOS and U2OS human OS cell lines from Biovector (Beijing, China) were grown in DMEM (Gibco, MA, USA) supplemented with 10% FBS (Gibco) and penicillin/streptomycin (Invitrogen, MA, USA) in a 5% CO 2 37°C incubator.

Techniques: Binding Assay, Luciferase, Cotransfection, Construct, Expressing, Transfection

Journal: Pharmacogenomics and Personalized Medicine

Article Title: Hsa_circ_0078767 Enhances Osteosarcoma Chemoresistance to Doxorubicin Through the Regulation of the miR-188-3p/GPX4 Axis

doi: 10.2147/PGPM.S473702

Figure Lengend Snippet: GPX4 is a miR-188-3p target ( A ) Overlap between predicted miR-188-3p and GPX4 interaction sites. ( B ) GPX4 levels in 65 and 61 non-respond and respond OS tissue samples, respectively, shown by qPCR. ( C ) Spearman correlations of miR-188-3p and GPX4 levels in DOX-resistant OS tissue samples. ( D and E ) Interaction between miR-188-3p and GPX4 shown by dual-luciferase reporter assays. ( F and G ) Interaction betweenmiR-188-3p and GPX4 shown by RIP. ( H ) GPX4 mRNA levels were analyzed in U2OS, U2OS/DOX, HOS, and HOS/DOX cells. ( I and J ) GPX4 levels were detected via qPCR in U2OS/DOX and HOS/DOX cells following miR-NC, miR-188-3p, anti-miR-NC, or anti-miR-188-3p transfection. *P < 0.05.

Article Snippet: The KHOS and U2OS human OS cell lines from Biovector (Beijing, China) were grown in DMEM (Gibco, MA, USA) supplemented with 10% FBS (Gibco) and penicillin/streptomycin (Invitrogen, MA, USA) in a 5% CO 2 37°C incubator.

Techniques: Luciferase, Transfection

Journal: Pharmacogenomics and Personalized Medicine

Article Title: Hsa_circ_0078767 Enhances Osteosarcoma Chemoresistance to Doxorubicin Through the Regulation of the miR-188-3p/GPX4 Axis

doi: 10.2147/PGPM.S473702

Figure Lengend Snippet: Hsa_circ_0078767 interacts with miR-188-3p to derepress the expression of GPX4. ( A ) Associations between hsa_circ_0078767 and GPX4 within DOX-resistant OS tissue samples were examined through Spearman correlation analyses. ( B and C ) GPX4 expression in U2OS/DOX and HOS/DOX cells following si-NC, si-hsa_circ_0078767, si-hsa_circ_0078767+anti-miR-NC, or si-hsa_circ_0078767+anti-miR-188-3p transfection, shown by qPCR and Western immunoblotting. *P<0.05.

Article Snippet: The KHOS and U2OS human OS cell lines from Biovector (Beijing, China) were grown in DMEM (Gibco, MA, USA) supplemented with 10% FBS (Gibco) and penicillin/streptomycin (Invitrogen, MA, USA) in a 5% CO 2 37°C incubator.

Techniques: Expressing, Transfection, Western Blot