Journal: Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan

Article Title: Modulation of expression of p16 and her2 in rat breast tissues of mammary hyperplasia model by external use of rupifang extract.

doi: 10.1016/s0254-6272(13)60087-4

Figure Lengend Snippet: Figure 2 her2 expression in rat breast tis- sues in each group (SP×200) A: bank control group (-); B: model group (+); C: the low dose group of Rupifang(+); D: the middle dose group of Rupifang (+); E: the high dose group of Rupifang (-).

Article Snippet: Rabbit-Anti-Human her2 and Rabbit-Anti-Human p16 SP immunohistochemical kit (Fuzhou Maixin Biological Technology Development Co., Ltd.); DAB chromogenic reagents (Beijing Zhongshan Gold Bridge Biotechnology Co., Ltd.); Estradiol Benzoate Injection: 1ml:1mg (Tianjin Jinyao Amino Acid Co., Ltd.); Progesterone injection: 1 ml∶20 mg (Tianjin Jinyao Amino Acid Co., Ltd.).

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: ERRF sensitizes ERBB2-positive breast cancer cells to lapatinib treatment likely by attenuating MCL1 and ERBB2 expression.

doi: 10.18632/oncotarget.16425

Figure Lengend Snippet: Figure 1: Lapatinib upregulates ERRF expression in SK-BR-3 and BT-474 breast cancer cell lines, and higher levels of ERRF correlate with lapatinib sensitivities and better patient survival. (A) Illustration of ERRF expression after lapatinib treatment (100 nM) for 12 and 24 hours in SK-BR-3 cells based on the data from the Array Express database [43]. (B, C) Lapatinib upregulates ERRF expression in BT-474 and SK-BR-3 breast cancer cell lines, both express ERRF and respond to ERBB2 inhibition. Lapatinib treatment was at the indicated concentrations (µM) for 48 hours, and ERRF expression was measured by real-time RT-PCR. (D) Row signal of ERRF in SK-BR3 and BT-474 breast cancer cell lines and lapatinib resistant clones derived from them, as detected in a microarray study in the GEO database [44]. (E) Correlation of ERRF mRNA levels with sensitivities to ERBB2 inhibition in breast cancer cell lines, according to published information [45] and the CCLE database [58]. (F) Higher levels of ERRF expression are associated with better prognosis in ERBB2-positive breast cancer, as determined in the BreastMark database (HR = 0.5442, p = 0.028, n = 107).

Article Snippet: Antibodies used in this study included: ERRF (1:1000 dilution, Sigma-Aldrich, St Louis, MO), ERBB2 (1:1000 dilution, OriGene, Rockville, MD), β-actin (1:5000 dilution, Sigma-Aldrich), and MCL1 (1:1000 dilution, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Inhibition, Quantitative RT-PCR, Clone Assay, Derivative Assay, Microarray

Journal: Oncotarget

Article Title: ERRF sensitizes ERBB2-positive breast cancer cells to lapatinib treatment likely by attenuating MCL1 and ERBB2 expression.

doi: 10.18632/oncotarget.16425

Figure Lengend Snippet: Figure 2: Ectopic expression of ERRF enhances lapatinib sensitivity in ERBB2-positive and lapatinib-resistant JIMT- 1 and MDA-MB-453 breast cancer cell lines. Lentivirus-mediated stable expression of ERRF was confirmed by western blotting (A, D) and cell viability was determined by the CCK-8 assay (B, E) and the sphere formation assay was in matrigel or in ultralow attachment plates (C, F). Lapatinib treatment was for 6 days at indicated concentrations in the CCK-8 assay and for 6 days at 2 µM in sphere formation assays. *p < 0.05; **p < 0.01; ***p < 0.001; pLHCX, vector control.

Article Snippet: Antibodies used in this study included: ERRF (1:1000 dilution, Sigma-Aldrich, St Louis, MO), ERBB2 (1:1000 dilution, OriGene, Rockville, MD), β-actin (1:5000 dilution, Sigma-Aldrich), and MCL1 (1:1000 dilution, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Western Blot, CCK-8 Assay, Tube Formation Assay, Plasmid Preparation, Control

Journal: Oncotarget

Article Title: ERRF sensitizes ERBB2-positive breast cancer cells to lapatinib treatment likely by attenuating MCL1 and ERBB2 expression.

doi: 10.18632/oncotarget.16425

Figure Lengend Snippet: Figure 3: Knockdown of ERRF decreases lapatinib sensitivities in ERBB2-positive breast cancer cells. SK-BR-3 (A–C) and BT-474 (D–I) cell lines were used, and the knockdown effect for siRNAs (#1 and #2) was confirmed by real-time PCR (A, F, H). The CCK-8 assay was used to detect cell viability (B, C, D, G, I), and sphere formation assay was also performed for BT-474 cells (E). Lapatinib treatments were at indicated times and doses (B, C, D). For BT-474 cells, different concentrations of siRNA were also applied (G, I) **p < 0.01; ***p < 0.001; C, negative control.

Article Snippet: Antibodies used in this study included: ERRF (1:1000 dilution, Sigma-Aldrich, St Louis, MO), ERBB2 (1:1000 dilution, OriGene, Rockville, MD), β-actin (1:5000 dilution, Sigma-Aldrich), and MCL1 (1:1000 dilution, Cell Signaling Technology, Danvers, MA).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, CCK-8 Assay, Tube Formation Assay, Negative Control

Journal: Oncotarget

Article Title: ERRF sensitizes ERBB2-positive breast cancer cells to lapatinib treatment likely by attenuating MCL1 and ERBB2 expression.

doi: 10.18632/oncotarget.16425

Figure Lengend Snippet: Figure 4: ERRF expression sensitizes xenograft breast cancer to the therapeutic effect of lapatinib and predicts response to lapatinib in breast cancer patients. Lapatinib resistant and ERBB2-positive JIMT-1 cells expressing ERRF or vector control (pLHCX) were injected subcutaneously into nude mice, lapatinib treatment was started at day 21, and tumor volumes (A) weights at excision (B) and images (C) are shown. (D). Detection of Ki67 in tumor sections by IHC staining. (E) Correlation of higher levels of ERRF expression with pathologic complete response (pCR) in patients treated with chemotherapy combined with lapatinib, as determined using a publically available data in the GEO database. NS, not significant; *P < 0.05; **P < 0.01.

Article Snippet: Antibodies used in this study included: ERRF (1:1000 dilution, Sigma-Aldrich, St Louis, MO), ERBB2 (1:1000 dilution, OriGene, Rockville, MD), β-actin (1:5000 dilution, Sigma-Aldrich), and MCL1 (1:1000 dilution, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Plasmid Preparation, Control, Injection, Immunohistochemistry

Journal: Oncotarget

Article Title: ERRF sensitizes ERBB2-positive breast cancer cells to lapatinib treatment likely by attenuating MCL1 and ERBB2 expression.

doi: 10.18632/oncotarget.16425

Figure Lengend Snippet: Figure 6: ERBB2 and MCL1 are involved in ERRF mediated sensitization to lapatinib treatment. (A) Expression of ERRF inversely correlates with that of MCL1 in breast cancer samples, as determined by the Pearson correlation analysis using data from the TCGA database (Supplementary Table 1). (B) Overexpression of ERRF inhibits ERBB2 and MCL1 expression in the JIMT-1 ERBB2 positive breast cancer cell line, as determined by western blotting. (C, D). Knockdown of ERRF in the BT-474 breast cancer cell line, as confirmed by real-time PCR (C), increases ERBB2 and MCL1 expression, as determined by western blotting (D). (E, F) Knockdown of MCL1 by siRNA transfection (50 nM) rescued decreased sensitivity of BT-474 cells to lapatinib mediated by ERRF knockdown (12.5 nM and 25 nM siRNA). ERRF and MCL1 expression levels were examined by real-time PCR or western blotting. Con, control; #1 and #2, siRNAs against ERRF.

Article Snippet: Antibodies used in this study included: ERRF (1:1000 dilution, Sigma-Aldrich, St Louis, MO), ERBB2 (1:1000 dilution, OriGene, Rockville, MD), β-actin (1:5000 dilution, Sigma-Aldrich), and MCL1 (1:1000 dilution, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Over Expression, Western Blot, Knockdown, Real-time Polymerase Chain Reaction, Transfection, Control