Breast Neoplasms ; chemistry ; genetics ; Chromogenic Compounds ; Female ; Genes, erbB-2 ; Humans ; In Situ Hybridization ; methods ; Receptor, ErbB-2 ; analysis

The gene types of breast cancer can be classified into three types according to its molecules: Luminal type A, Luminal type B, HER-2-positive type, triple negative type. Authors combined pathological characteristics of breast cancer, biological characteristics, and comprehensive treatment, used syndrome typing based medication, and explored treatment meticulous ideas and methods of "treating the same disease with different methods" as well as "different treatment methods in accordance with patients individually".
Biomarkers, Tumor ; genetics ; Breast Neoplasms ; classification ; genetics ; therapy ; Female ; Humans ; Receptor, ErbB-2 ; genetics

Human epidermal growth factor receptor 2 (HER-2) plays an important role in carcinogenesis and development of urothelial carcinoma. Overexpression of HER-2 is associated with poor prognosis of urothelial carcinoma. Although there is no significant benefit from anti-HER-2 targeted therapies of monoclonal antibody and tyrosine kinase inhibitor, Anti-HER-2 antibody-drug conjugate (HER-2-ADC) has shown a promising efficacy in urothelial carcinoma patients with HER-2 overexpression. Therefore, effectively screening the potential beneficiaries of HER-2-ADC drugs has become a new challenge. However, standardized HER-2 scoring system for urothelial carcinoma has yet to be developed. Thus, the Committees organized experts to reach this expert consensus based on the clinical practice of HER-2 expression, gene amplification and mutation testing in urothelial carcinoma, combined with the current research progress and internal discussion of committee members, in order to construct HER-2 testing standard of urothelial carcinoma and improve the accuracy of interpretation, to guide the clinical application.
Carcinoma, Transitional Cell/genetics* ; Consensus ; Humans ; Receptor, ErbB-2/genetics* ; Urinary Bladder Neoplasms/genetics*

Breast cancer is the most common malignant tumor in women, of which the majority is early breast cancer (EBC). The strategy of postoperative adjuvant treatment relies mainly on the clinicopathologic characteristics of patients, but there are certain deficiencies if only depending on it to assess treatment benefits and disease prognosis. Multigene testing tools can evaluate the prognosis and predict therapeutic effects of breast cancer patients to guide the clinical decision-making on whether to use adjuvant chemotherapy, radiotherapy, and endocrine therapy by detecting the expression levels of specific genes. The consensus-writing expert group, based on the characteristics, validation results, and accessibility of the multigene testing tools and combined with clinical practice, described the result interpretation and clinical application of OncotypeDx(®) (21-gene), Mammaprint(®) (70-gene), RecurIndex(®) (28-gene), EndoPredict(®)(12-gene), and BreastCancerIndex(®) (BCI, 7-gene) for hormone receptor-positive and human epidermal growth factor receptor 2-negative EBC. The development and validation process of each tool was also briefly introduced. It is expected that the consensus will help guide and standardize the clinical use of multigene testing tools and further improve the level of precise treatment for EBC.
Humans ; Female ; Breast Neoplasms/genetics* ; Consensus ; East Asian People ; Prognosis ; Chemotherapy, Adjuvant ; Receptor, ErbB-2/genetics*

To purify the extracellular domain of HER2 in vitro and improve its prokaryotic expression abundance, the cDNA fragment encoding extracellular domain of HER2 was obtained by PCR and cloned into the expression vector pGEX-6P-1. After transforming it into Escherichia coli BL21, we instituted an investigation of different inducing conditions to try out the optimal condition for expressing soluble fusion protein. As for insoluble inclusion bodies, they were dissolved in 8 M Urea and refolded in refolding buffer. The soluble protein and the refolded protein were purified with Glutathione Sepharose 4B, respectively. The results showed that both the soluble and insoluble protein existed in Escherichia coli, but the majority was insoluble. It is beneficial to the expression of soluble fusion protein by induction at lower temperature (30 degrees C) and higher optical density (A600= 1.8) with the use of certain additive in medium. By purification of the supernatant of the lysate and refolded protein, the yield of the fusion protein was about 1.23 mg per liter culture. As a result, we have obtained the maximum soluble extracellular domain of HER2 protein, and thus have laid a foundation for further work on functional study and antibody preparation for HER2.
Cloning, Molecular ; Escherichia coli ; genetics ; Genes, erbB-2 ; genetics ; Humans ; Prokaryotic Cells ; metabolism ; Protein Folding ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Receptor, ErbB-3 ; biosynthesis ; genetics

OBJECTIVETo construct a plasmid carrying small interfering RNA (siRNA) targeting human C-erbB2 gene (pGenesil- erbB2) and test its effect on Her-2 expression at the post-transcriptional level in human colon cancer cell lines HT-29 cells that highly express erbB2.

METHODSA HT-29 cell line that highly expressed CerbB-2 was selected using immunohistochemical method. The double-stranded siRNA targeting human CerbB-2 cDNA and the negative control fragment were cloned into pGenesil-1 vector, and after identification and sequence analysis, the constructed pGenesil-erbB2 plasmid was transfected into the selected HT-29 cell line.

RESULTSThe pGenesil-erbB2 plasmid was successfully constructed and stably transferred into HT-29 cells. The transfection resulted in significant inhibition of Her-2 protein expression in the HT-29 cells, as shown by Western blotting.

CONCLUSIONThe pGenesil-erbB plasmid we constructed can be stably transfected into HT-29 cells to inhibit the expression of Her-2 protein, and can be useful in further studies of increasing the radiosensitivity of HT-29 cell lines.

Base Sequence ; Genes, erbB-2 ; genetics ; HT29 Cells ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; RNA Interference ; RNA, Small Interfering ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Transfection

OBJECTIVETo construct human phage single-chain antibody (scFv) library against breast cancer, and to identify anti-HER2 specific antibodies from the human phage display scFv library to offer a stronger affinity sequence targeting HER2 for fusion protein targeting HER2 and CXCR4.

METHODSTotal RNA was extracted from the adjacent lymphatic tissue harvested from breast cancer patients. The variable regions of the whole antibody were amplified by using RT-PCR and were cloned into the vector pCANTAB-5E through a linker. The products were electroporated into competent E.coli TG1 cells. Recombinant phages specific for breast cancer cells were enriched in SKBR-3 after four rounds. The antigen-positive clones were selected by ELISA and immunohistochemistry.

RESULTSThe fragment of VH and VL were about 375 and 330 bp and were linked in vitro to form scFv of 750 bp that was resistant to the breast cancer HER2 single strand. A fusion phage display library that contained total of 2.48×10(8) pfu /ml was established. ELISA and immunohistochemical results confirmed that the antibody has a strong affinity with HER2 antigen in breast cancer tissue. Compared to human IgG antibody, a scFv phage library against human breast cancer was successfully constructed with high capacity. The scFv was highly specific to HER2 antigen and the sequencing results indicated that VL and VH genes were highly homologous with the variable region of human antibody.

CONCLUSIONThis strategy may achieve new targeted antibody resistant to the breast cancer for clinical treatment and provide a carrier that uses HER2 as a target of the fusion protein for anti-tumor therapy.

Breast Neoplasms ; genetics ; immunology ; Female ; Humans ; Peptide Library ; Receptor, ErbB-2 ; immunology ; Single-Chain Antibodies ; immunology

Breast Neoplasms/genetics* ; Female ; Gene Amplification ; Humans ; In Situ Hybridization, Fluorescence ; Receptor, ErbB-2/metabolism*

Humans ; Female ; Breast Neoplasms/pathology* ; Multiomics ; Trastuzumab ; Receptor, ErbB-2/genetics* ; Machine Learning