Journal: Frontiers in Genetics

Article Title: Analysis of gene expression patterns modulated by tuberculous pleural effusion–derived exosomal miRNAs in lung cancer

doi: 10.3389/fgene.2026.1828734

Figure Lengend Snippet: qRT-PCR validation of target gene expression following transfection with candidate miRNA mimics or inhibitors. A549 cells were transfected with sequence-specific miRNA mimics or inhibitors, with a non-targeting RNA serving as the control. Target gene expression was quantified by qRT-PCR at the indicated time points. (A) Expression of NDUFA2 , NDUFA4 , NDUFB1 , NDUFB4 , RPS15A , RPS21 , RPS25 , and RPS27 24 h after transfection with an hsa-miR-1290 mimic or inhibitor. (B) Expression of COX7A2 3 h after transfection with an hsa-miR-224 mimic or inhibitor. (C) Expression of RHOA , PRKACB , and RPS27 24 h after transfection with an hsa-miR-185 mimic or inhibitor. (D) Expression of RPS15A , RPS21 , RPS25 , and RPS27 12 h after transfection with an hsa-miR-30a mimic or inhibitor. All experiments were performed in triplicate. Data are presented as mean ± standard error of the mean (n = 3). P < 0.05 vs. control unless otherwise indicated. qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: To investigate the correlation between miRNAs from TPE-derived exosomes and gene expression in lung cancer cells, A549 cells were transfected with miRNA mimics or inhibitors for hsa-miR-1290, hsa-miR-185, hsa-miR-30a, and hsa-miR-224 (200 pM) (Bioneer, Daejeon, Republic of Korea).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Targeted Gene Expression, Transfection, Sequencing, Control, Expressing, Real-time Polymerase Chain Reaction

Journal: Breast Cancer : Targets and Therapy

Article Title: Hsa_circ_0000520 Promotes Invasion and Metastasis of Breast Cancer Cells by Targeting HSP90AA1

doi: 10.2147/BCTT.S600770

Figure Lengend Snippet: Hsa_circ_0000520 is verified and characterized in BC cells. ( A ) Analysis of the GSE101123 microarray through bioinformatic approaches revealed a set of expressed circRNAs, among which hsa_circRNA_0000520 was detected. ( B ) Expression levels of hsa_circ_0000520 in BC specimens were analyzed using data from GSE101123 . ( C ) Sanger sequencing confirmed the sequence of this circRNA. ( D ) Detect the presence of hsa_circRNA_0000520 from cDNA and gDNA in HS578T cells by RT-PCR. Divergent primers successfully amplified hsa_circRNA_0000520 only from cDNA. ( E ) The expression profiles of hsa_circ_0000520 and its linear counterpart RPPH1 were examined after RNase R treatment. ( F ) RNA levels of hsa_circ_0000520 and RPPH1 were quantified in HS578T cells exposed to actinomycin D at the specified time point. ( G ) Hsa_circ_0000520 was predominantly distributed in the cytoplasm of BC cells, with U6 and GAPDH used as nuclear and cytoplasmic controls, respectively. ( H ) RT-qPCR assay measured hsa_circ_0000520 expression in 42 pairs tumor and neighboring tissues. ( I ) RT-qPCR assay measured hsa_circ_0000520 expression in various BC cell line. * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: Lentiviruses for overexpression and knockdown of hsa_circ_0000520, as well as negative control, were procured from Genechem, Shanghai, China.

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

Journal: Breast Cancer : Targets and Therapy

Article Title: Hsa_circ_0000520 Promotes Invasion and Metastasis of Breast Cancer Cells by Targeting HSP90AA1

doi: 10.2147/BCTT.S600770

Figure Lengend Snippet: BC patients show increased serum expression of hsa_circ_0000520. ( A ) The standard curve was employed for the purpose of absolute quantitation. ( B ) The serum expression level of hsa_circ_0000520 from healthy people (n = 100), individuals with benign breast disease (n=100), and BC (n = 128) were quantified using RT-qPCR. ( C ) The abundance of hsa_circ_0000520 in different stages of the disease. * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: Lentiviruses for overexpression and knockdown of hsa_circ_0000520, as well as negative control, were procured from Genechem, Shanghai, China.

Techniques: Expressing, Quantitation Assay, Quantitative RT-PCR

Journal: Breast Cancer : Targets and Therapy

Article Title: Hsa_circ_0000520 Promotes Invasion and Metastasis of Breast Cancer Cells by Targeting HSP90AA1

doi: 10.2147/BCTT.S600770

Figure Lengend Snippet: Hsa_circ_0000520 increases proliferation, migration and invasion of BC in vitro. ( A ) Transfection efficiency of the overexpressed hsa_circ_0000520 lentivirus in MDA-MB-231 cells was assessed by RT-qPCR. ( B ) Transfection efficiency of hsa_circ_0000520 lentivirus knockdown in MDA-MB-231 cells was analyzed by RT-qPCR. ( C and D ) Cell growth in the distinct MDA-MB-231 groups was quantified through CCK-8 assays. ( E and F ) The growth of BC cells was measured by colony formation assay. ( G and H ) Scratch wound analysis was used to evaluate changes in MDA-MB-231 cell migration following hsa_circ_0000520 overexpression or knockdown. ( I and J ) Evaluation of how hsa_circ_0000520 overexpression and knockdown influence the migratory capacity of MDA-MB-231 cells was conducted using a migration assay (200x). ( K and L ) Evaluation of how hsa_circ_0000520 overexpression and knockdown influence the invasive behavior of MDA-MB-231 cells was assessed using an invasion assay (200x). * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001.

Article Snippet: Lentiviruses for overexpression and knockdown of hsa_circ_0000520, as well as negative control, were procured from Genechem, Shanghai, China.

Techniques: Migration, In Vitro, Transfection, Quantitative RT-PCR, Knockdown, CCK-8 Assay, Colony Assay, Over Expression, Invasion Assay

Journal: Breast Cancer : Targets and Therapy

Article Title: Hsa_circ_0000520 Promotes Invasion and Metastasis of Breast Cancer Cells by Targeting HSP90AA1

doi: 10.2147/BCTT.S600770

Figure Lengend Snippet: Hsa_circ_0000520 can bind to HSP90AA1. ( A and B ) ChIRP assay was conducted with the biotin-tagged hsa_circ_0000520 probe in MDA-MB-231 cell lysates, with subsequent silver staining, and the protein bands were further examined using MS. ( C ) WB analysis of the RNA pulldown material confirmed that HSP90AA1 was captured by hsa_circ_0000520. ( D ) The association between HSP90AA1 and hsa_circ_0000520 is further demonstrated by RIP reverse analysis. *** P <0.001.

Article Snippet: Lentiviruses for overexpression and knockdown of hsa_circ_0000520, as well as negative control, were procured from Genechem, Shanghai, China.

Techniques: Silver Staining

Journal: Breast Cancer : Targets and Therapy

Article Title: Hsa_circ_0000520 Promotes Invasion and Metastasis of Breast Cancer Cells by Targeting HSP90AA1

doi: 10.2147/BCTT.S600770

Figure Lengend Snippet: Tanespimycin inhibited the invasion and migration of BC cells. ( A ) After treatment with the HSP90AA1 inhibitor Tanespimycin, the WB experiment verified that HSP90AA1 expression was decreased in BC cells. ( B and C ) The CCK-8 assays were employed to determine the viability of MDA-MB-231 cells with hsa_circ_0000520 overexpression or knockdown, treated with or without 1.79 μM Tanespimycin for 48 hours. ( D and E ) Colony formation assays were performed to assess MDA-MB-231 proliferation with hsa_circ_0000520 overexpression or knockdown, treated with or without 1.79 μM Tanespimycin for 48 hours. ( F and G ) scratch assays were performed on BC cells with hsa_circ_0000520 overexpressed or knocked down, treated with or without 1.79 μM Tanespimycin for 48 hours. ( H and I ) The effect of hsa_circ_0000520 overexpression or knockdown, with or without 1.79 μM Tanespimycin treatment for 48 hours, on MDA-MB-231 cell migration was assessed by the migration assays (200x). ( J and K ) The effect of hsa_circ_0000520 overexpression or knockdown, treated with or without 1.79 μM Tanespimycin for 48 hours, on MDA-MB-231 cell invasion was assessed using invasion assays (200x). β-actin served as an internal reference. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Lentiviruses for overexpression and knockdown of hsa_circ_0000520, as well as negative control, were procured from Genechem, Shanghai, China.

Techniques: Migration, Expressing, CCK-8 Assay, Over Expression, Knockdown