Comparison of fluorescence in-situ hybridisation with dual-colour in-situ hybridisation for assessment of HER2 gene amplification of breast cancer in Hong Kong

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The miR-143 target TARDBP may serve as a marker for cervical cancer

Conference Theme: Translating Science into MedicineBACKGROUND: The microRNA miR-143 has been demonstrated to be downregulated in cervical cancer. However its targets in cervical cancer are not well characterized but could potentially yield useful markers for the malignancy. OBJECTIVES: (1) To determine the cellular expression pattern of miR-143 in normal and cancerous cervices. (2) To explore miR-143 and its targets as makers of cervical cancer. DESIGN: Cells expressing miR-143 in normal cervix, CIN, and SCC were identified with in situ hybridization. Ten potential miR-143 target genes were chosen from in silico prediction using three algorithms (PicTar, miRanda and miRtarget2). Their transcript and protein expression levels were characterized in five cervical cancer cell lines (HeLa, SiHa, CaSki, C4-I and C33A) and in HeLa with miR-143 overexpression. One of the genes was verified as true target by luciferase reporter assay. Its expression in cervical tissues was assessed with immunohistochemistry. RESULTS: miR-143 was strongly expressed in blood vessels and smooth muscle in all samples. Weaker and scattered staining could be detected in normal epithelium. However miR-143 could not be found in SCC cells. Ten potential miR-143 target genes were chosen from in silico prediction (ITGA6, MSI2, PHF6, TARDBP, VAPB, MAP3K7, CRELD1, AKAP6, CBFB, RBM24). All candidates except MAP3K7 displayed certain degree of upregulation in at least one of the cervical cancer cell lines in comparison with HaCat. However, only TARDBP and PHF6 were downregulated by miR-143 overexpression in HeLa. Luciferase transcript with the 3’UTR of TARDBP was also found to be downregulated by miR-143. Such down-regulation was abolished when the miR-143 binding site was mutated. TARDBP was found to be overexpressed in SCC in comparison with CIN2/3, CIN1 and normal cervix by immunocytochemistry. CONCLUSION: TARDBP is a target of miR-143 and is overexpressed in SCC. TARDBP may serve as an immune-histochemical marker for detection of cervical cancer

Potential targets in miRNA 143 in cervical cancers

Congress Theme: Translating Discoveries into Prevention and Cure

Comparison of fluorescence in-situ hybridisation with dual-colour in-situ hybridisation for assessment of HER2 gene amplification of breast cancer in Hong Kong

OBJECTIVES: To compare the PathVysion fluorescence in-situ hybridisation assay with the INFORM HER2 Dual in-situ hybridisation assay on 104 invasive breast cancers with a broad spectrum of immunohistochemistry scores. METHODS: This case series involved consecutive patients diagnosed with invasive breast carcinoma with equivocal immunohistochemistry score and referred for further HER2 assessment from the departments of Surgery and/or Clinical Oncology of the two hospitals between January 2013 and February 2014. An additional 10 cases with negative HER2 immunohistochemistry and 11 cases with positive HER2 immunohistochemistry were further included. RESULTS: The results of both fluorescence in-situ hybridisation and dual in-situ hybridisation were available in 99 of 104 cases, respectively. Student'st test showed no statistically significant difference in the mean number of HER2 count, CEP17 copies, or HER2/CEP17 ratio between that obtained by fluorescence in-situ hybridisation and that obtained by dual in-situ hybridisation. Pearson's correlation of results for the two assays was strong for HER2/CEP17 signal ratio (R=0.963, P<0.001) and mean HER2 copies per nucleus (R=0.897, P<0.001). Overall agreement was 96.0% (95 out of 99 cases, k0.882). Three of the four discordant cases were equivocal for either fluorescence in-situ hybridisation or dual in-situ hybridisation. The results of immunohistochemistry 0/1+ and 3+ cases showed 100% concordance between the two assays. The failure rate was 0.96% for fluorescence in-situ hybridisation and 3.85% for dual in-situ hybridisation. Cases that failed for fluorescence in-situ hybridisation were successful for dual in-situ hybridisation and vice versa. CONCLUSIONS: Our study showed that dual in-situ hybridisation is a reliable and useful option for HER2 testing in breast cancer

Comparison of the genoflow human papillomavirus (HPV) test and the linear array assay for HPV screening in an Asian population

High-risk human papillomavirus (HR-HPV) DNA detection in cervical cytology samples is useful for primary screening of cervical cancer and for triage of patients with equivocal cytological findings. The GenoFlow HPV array test (GF assay; Diagcor Bioscience Inc., Hong Kong) was recently developed to detect 33 HPV genotypes by a "flowthrough" hybridization technology. In this study, we assessed the analytical sensitivity and reproducibility of the GF assay and compared its genotyping results with those of the Linear Array (LA) assay (Roche Molecular Diagnostics, Indianapolis, IN), using 400 archived liquid-based cytology samples representing the full range of cytology findings. Genotyping findings of the GF and LA assays were concordant or compatible for 93.44% of tested samples, with a good ( κ = 0.797) to very good ( κ 0.812) strength of agreement for assay-common and oncogenic HPV types, respectively. The two assays showed good ( κ = 0.635) agreement in detecting infections with multiple HPV genotypes. The lowest detection limits of the GF assay for HPV16 and HPV18 were 25 copies and 20 copies, respectively. Repeat testing of 60 samples by use of two different lots of the GF assay revealed no discordant results, suggesting good reproducibility of the assay. Both assays achieved approximately 80% and 100% sensitivity for identifying cases of atypical squamous cells of undetermined significance (ASC-US) and low-grade squamous intraepithelial lesions (LSIL) with subsequent detection of LSIL+ and high-grade squamous intraepithelial lesions or higher (HSIL+) in 2 years, respectively. Among ASC-US samples, the GF assay achieved the highest specificity (23.08%) for indicating subsequent identification of HSIL compared with the LA (19.23%) and Hybrid Capture 2 (HC2) (8.97%) assays. The GF and LA assays showed significant discrepancy in detecting HPV genotypes 11, 26, 39, 52, and 66. More sensitive detection of HPV52 by GF assay offers an advantage in regions where HPV52 is more prevalent. The sensitivity of the GF assay for detecting patients with HSIL+ was noninferior to that of the LA assay. Copyright © 2012, American Society for Microbiology. All Rights Reserved.link_to_OA_fulltex