Journal: Cancers

Article Title: ANKRD1 Promotes Breast Cancer Metastasis by Activating NF- κ B-MAGE-A6 Pathway

doi: 10.3390/cancers16193306

Figure Lengend Snippet: High ANKRD1 expression correlates with poor prognosis, aggressiveness and tumor progression of breast cancer. ( A ) Kaplan–Meier survival analysis of distant metastasis-free survival (DMFS) breast cancer patients with metastatic disease based on ANKRD1 expression using mRNA gene chip (HR; the hazard ratio) ( https://kmplot.com ). The following parameters were set: (1) lymph node status: positive; (2) probe set: only JetSet best probe; (3) grade: 3; and (4) survival: DMFS. Patients with higher ANKRD1 expression showed lower survival rates compared to those with low expression of ANKRD1. ( B , C ) Real-time PCR and Western blot results showing ANKRD1 fold change in normal breast (MCF-10A) and non-metastatic (ZR-75-30, T47D and MCF-7) and highly metastatic breast cancer cells (LM-2 and MDA-MB-231). ( D ) IHC staining (left panel) of ANKRD1 in human breast tissue array showing progressively higher ANKRD1 expression depending on the grade of breast cancer tissue. The right panel shows H&E-stained sections.

Article Snippet: A human breast cancer tissue array containing 100 cancer cases of different grades was purchased (BR10011, US Biomax, Rockville, MD, USA), and mouse organs were collected and maintained in 4% paraformaldehyde solution and used to detect ANKRD1 expression.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Staining

Journal: Cancers

Article Title: ANKRD1 Promotes Breast Cancer Metastasis by Activating NF- κ B-MAGE-A6 Pathway

doi: 10.3390/cancers16193306

Figure Lengend Snippet: ANKRD1 knockdown reduces metastasis efficiency of breast cancer cells in two in vivo metastasis models. ( A , B ) Representative bioluminescence (BLI) images of animals showing primary tumors (PT) at day 21, derived from orthotopic injections of LM-2 vector control (vector) and LM-2 ANKRD1 knockdown (LM-2 sh14) cells into the mammary fat pad. On day 21 after orthotopic injection into the mammary fat pad, tumor growth at the injection site and GFP was detected by IVIS and quantified based on radiant efficiency, comparing vector and LM-2 sh14 groups (day 21, n = 10 per group). ( C ) Comparison of the size of primary tumors from vector and LM-2 sh14 groups after removing the primary tumor. ( D ) Quantitation of tumor volume (cm 3 ) comparing control and LM-2 sh14 groups. The results show no significant change in tumor volume and tumorigenesis in the LM-2 sh14 cell group compared to the vector. Data presented as mean ± SD. ( E ) Radiant efficiency of the lung and liver comparing vector and LM-2 sh14 groups and ex vivo GFP signals, detected using IVIS. ( F , G ) Representative BLI images of animals on the first day of tail vein injection and quantitation of radiant efficiency in mice (day 1, n = 10 per group). ( H ) Liver metastasis with control and LM-2 sh14 cells. The results show increased metastasis of cancer cells in control compared to LM-2 sh14 group. ( I , J ) H&E staining of liver tissue from tail-vein-injected mice; red circles show the cancerous lesions in the tissues, which are increased in the control group compared to the LM-2 sh14 group. LM-2 Vector (control): LM-2 with empty vector; LM-2 sh14: LM-2-ANKRD1 knockdown with sh14. Ten NOD.SCID mice were used for each set, and all experiments were performed in triplicate.

Article Snippet: A human breast cancer tissue array containing 100 cancer cases of different grades was purchased (BR10011, US Biomax, Rockville, MD, USA), and mouse organs were collected and maintained in 4% paraformaldehyde solution and used to detect ANKRD1 expression.

Techniques: Knockdown, In Vivo, Derivative Assay, Plasmid Preparation, Control, Injection, Comparison, Quantitation Assay, Ex Vivo, Staining

Journal: PLoS ONE

Article Title: RecQL4 Helicase Amplification Is Involved in Human Breast Tumorigenesis

doi: 10.1371/journal.pone.0069600

Figure Lengend Snippet: (A) Western blot detection of RecQL4 protein level in HMEC, MCF-10F and five breast tumor cell lines. β-Actin was used to verify equal loading of proteins. (B) Analysis of RecQL4 expression by quantitative real time PCR in normal breast tissues and breast cancer specimens with different pathological grades. TissueScan breast cancer tissue qPCR array was purchased from Origene. RecQL4 expression detected in normal breast tissues was considered as 1. The data represent mean ± SD from three independent experiments.

Article Snippet: To verify whether or not RecQL4 expression is elevated in human breast tumor tissue samples, RecQL4 mRNA level was examined using TissueScan Human Breast cancer tissue qPCR array (Origene, Rockville, MD 20850).

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction

Journal: Cancers

Article Title: miR-302 Suppresses the Proliferation, Migration, and Invasion of Breast Cancer Cells by Downregulating ATAD2

doi: 10.3390/cancers14184345

Figure Lengend Snippet: The expression of ATAD2 is elevated in breast cancer tissues and cell lines. ( A ) RT-PCR was performed to analyze the mRNA expression of ATAD2 in 33 paired samples of the non-tumor breast (N) and tumor breast (T) tissues. β-actin was used as an internal control ( B ) The relative expression level of ATAD2 mRNA was identified by qRT-PCR analysis in 15 paired samples of the non-tumor breast (N) and tumor breast (T) tissues. ( C ) ATAD2 expression in TCGA cohort using the GEPIA ( http://gepia.cancer-pku.cn/detail.php , accessed on 5 September 2017). ( D ) ATAD2 expression in normal breast ( n = 10) and breast cancer tissues including stage IIA ( n = 12), stage IIB ( n = 8), stage IIIA ( n = 12), stage IIIB ( n = 4), and stage IIIC ( n = 14). Original magnification, 100×. The histogram shows the relative intensity of ATAD2-positive cells. The mean values and the standard error were obtained from two individual experiments. ** p < 0.01, # p < 0.001. ( E ) The level of ATAD2 protein and mRNA in human breast cancer cell lines was verified via Western blot and RT-PCR analysis, respectively. Relative intensities were measured by ImageJ. Uncropped blots can be found in .

Article Snippet: A human breast cancer tissue array (60 cases; CBA) was purchased from SuperBioChips (Seoul, Korea).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot