• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.3903-3909
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• 2016

Purpose: To evaluate the expression of Her2/neu and Ki-67 in benign and malignant gallbladder lesions, and to establish correlations with clinico-pathologic parameters. Materials and Methods: A retrospective analysis was conducted on formalin fixed paraffin embedded (FFPE) benign (n=25) and malignant gallbladder (n=25) tissue samples. Hematoxylin and eosin stained slides of each case were reviewed for: type of malignancy (whether adenocarcinoma, squamous cell carcinoma, or any other type), grade (well, moderate, and poor), depth of invasion, pre-neoplastic changes in adjacent mucosal epithelium like metaplasia and dysplasia. Immunohistochemistry for Her 2 neu and Ki-67 was performed and data analysis was conducted using SPSS 17 software. Chi-square test was used to compare categorical/dichotomous variables. P value of ${\leq}0.05$ was considered significant. Results: The difference of Her 2 neu expression and Ki67 index between benign and malignant groups was found to be statistically significant. Her2/neu positivity did not have any significant correlation with various clinicopathological parameters other than liver involvement. 5 cases of gallbladder cancer showed both Her2/neu and Ki67 positivity. Ten cases were Ki67 positive but Her2/neu negative while one case was Her2/neu positive but Ki67 negative. Conclusions: The present study demonstrated overexpression of Her2/neu and Ki67 in gallbladder cancer. A trend of decreasing Her2/neu expression with increasing grade of tumor was observed. Furthermore, greater Ki67 positivity was found in cases with lymph node metastasis and distant metastasis. Future studies with a larger number of patients will be required to precisely define the correlation of Her2/neu expression and Ki67 positivity with clinicopathological parameters. The results however are encouraging and suggest evaluation of Her2/neu as a candidate for targeted therapy.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.1023-1026
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• 2013

Cervical cancer is the most important female gynecological cancer, the second leading cause of cancer mortality in women worldwide and the second most common cancer in Thai women. The major cause of cervical cancer is persistent infection of human papillomavirus (HPV), leading to abnormal epithelial lesions, with progression to precancerous and invasive cancer. This study was conducted to investigate the frequency and type distribution of HPV in Thai women who had abnormal cytology. HPV detection from FFPE confirmed abnormal of high grade cervical intraepithelial lesions were for SPF-10-Innogenic Line Probe Assay. HPV-positivity was detected in 320/355 cases (90.14%) and HPV-negativity in 35/355 (9.86%). HPV-positive was found 147/320 cases (41.4%) of single infection, whereas 173/320 cases (48.7%) showed the multiple HPV infection. The most common seven types were HPV-16, -52, -18, -11, -51, -31 and -33, in that order. HPV 16 and 18, the important oncogenic HPV type, were observed in 64.8% of HSIL cases. Interestingly, a high proportion of multiple infections was found in this study and more than ten types could be detected in one case. Therefore, HPV infection screening program in women is essential, particularly in Thailand. Effective primary and secondary prevention campaigns that reinforce HPV screening for HPV detection and typing may be decrease the incidence and mortality of cervical cancer in the future and may lead to significantly improve the quality of life in Thai women.

• 대한임상검사과학회지
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• 제45권2호
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• pp.66-72
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• 2013

Fine Needle Aspiration Biopsy Cytology (FNABC), which is known as the most accurate and cost-effective method for diagnosis of the thyroid nodule, may still result in indeterminate cases that are cellular paucity and show minor nuclear atypia. However, most cases are associated with suspicion of papillary thyroid carcinoma (PTC). A B-type Raf kinase (BRAF) mutation was found in about half of PTCs which is currently helping us to differentiate malignancies from benign lesions. Cases studied included 46 histological, confirmed PTC cases. FNABC 102 cell paucity and 74 atypia benign cases were previously diagnosed as suspicious of PTC using cytologic examination. These cases were analyzed for BRAF mutation by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with a new restriction enzyme. In this study, the sensitivity and specificity were calculated and, BRAF mutation was detected by means of a histological method in 23 of 46 cases of PTC and no mutation was found in 22 cases. However, one case was not detected. In using FNABC, BRAF mutation was detected in 6 of 102 cases in cell paucity and in 11 of 74 cases in the atypia. Two cases were not detected in the atypia. The sensitivity and specificity of PCR-RFLP in FNABC were 60% and 97.4% respectively. Assessment of Formalin Fixed Paraffin Embedding (FFPE) block demonstrated similarly a 51.1% positive and 48.9% negative in PTC. Evaluation of BRAF mutation revealed high specificity and low sensitivity in using FNABC method. This study suggests that BRAF mutation analysis should be useful for the clinical diagnosis of PTC in FNABC with cytological findings suspicious for PTC.

• Asian Pacific Journal of Cancer Prevention
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• 제17권12호
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• pp.5229-5235
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• 2016

Prostate cancer (PCa) remains one of the most widespread and perplexing of all human malignancies. Assessment of gene expression is thought to have an important impact on cancer diagnosis, prognosis and therapeutic decisions. In this context, we explored combined expression of PCa related target genes AMACR and PCA3 in 126 formalin fixed paraffin embedded prostate tissues (FFPE) from Moroccan patients, using quantitative real time reverse transcription-PCR (RT-qPCR). This quantification required data normalization accomplished using stably expressed reference genes (RGs). A panel of twelve RG was assessed, data being analyzed using GenEx V6 based on geNorm, NormFinder and statistical methods. Accordingly, the hnRNP A1 gene was identified and selected as the most stably expressed RG for reliable and accurate gene expression quantification in prostate tissues. The ratios of both PCA3 and AMACR gene expression relative to that of the hnRNP A1 gene were calculated and the performance of each target gene for PCa diagnosis was evaluated using receiver-operating characteristics. PCA3 and AMACR mRNA quantification based on RT-qPCR may prove useful in PCa diagnosis. Of particular interesting, combining PCA3 and AMACR quantification improved PCa prediction by increasing sensitivity with retention of good specificity.

• 대한의생명과학회지
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• 제29권1호
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• pp.48-52
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• 2023

Formaldehyde use is associated with serious health risks, which can affect medical personnel and technicians. Therefore, we investigated the efficacy of an alternative fixative, with respect to two types of formalin fixatives, by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, immunohistochemical (IHC) staining, and RNA extraction. For H&E staining, the circular nucleus was stained dark blue by the basic dye hematoxylin and the cytoplasm was stained red by the acid dye eosin in all three fixative samples. No difference was found in the Duksan General Science (DGS), Sigma-Aldrich, and Core-Fix fixative samples (Corebiotech) used to fix kidney tissue, after PAS staining. IHC staining showed that CD4 was significantly increased in the lippolysaccharide (LPS)-treated group compared to the control group (vehicle), confirming the changes in specific molecules. The quantity and quality of RNA from tissues fixed in the three types of fixatives were evaluated. The average concentration of RNA was 106 ng/µL and average purity at A 260/280 ratio was 1.7~2.0, regardless of fixative used. For quality of protein, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was confirmed by Western blotting. In conclusion, Core-Fix can be used as a fixative for pathological tissues, in histological and molecular diagnoses.

• 대한임상검사과학회지
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• 제49권2호
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• pp.79-87
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• 2017

GenesWell$^{TM}$ BCT는 호르몬 수용체 양성, HER2 음성, 및 pN0 또는 1인 조기 유방암 환자의 10년내 타 장기 전이 재발 위험도를 제시하는 다지표 예후 예측 검사로, 예후에 대한 위험을 BCT Score로 제시한다. 본 연구에서는 GenesWell$^{TM}$ BCT의 분석적 성능을 검사하였다. 조기 유방암 환자의 FFPE 검체로 부터 추출한 RNA를 대상으로 GenesWell$^{TM}$ BCT 수행하여, 12개 유전자의 발현값을 측정하였다. GenesWell$^{TM}$ BCT의 최소검출한계, 공란 한계 및 측정 범위는 단계 희석한 RNA 검체를 사용하여 평가하였으며, 분석적 정밀도 및 특이도 시험은 BCT Score에 따라 저위험군, 고위험군 그리고 경계선 주변으로 나누어진 3개의 RNA 검체를 이용하여 시험하였다. GenesWell$^{TM}$ BCT는 $1ng/{\mu}L$ 미만의 RNA 검체에서 RNA를 측정할 수 있었으며, 다기관에서 수행된 분석적 정밀도 시험에서 반복성 100% 및 재현성 98.3%의 결과를 확인할 수 있었다. 또한, 분석적 특이도 시험을 통해, 간섭 물질이 검체의 재발 위험성 판정에 영향을 미치지 않음을 확인할 수 있었다. 이들 결과는 GenesWell$^{TM}$ BCT가 95% 이상의 항상성을 나타내는 높은 분석적 성능을 가지고 있음을 제시한다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.7071-7076
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• 2015

Epidermal growth factor receptor (EGFR) is one of the targeted molecular markers in many cancers including lung malignancies. Gefitinib and erlotinib are two available therapeutics that act as specific inhibitors of tyrosine kinase (TK) domains. We performed a case-control study with formalin-fixed paraffin-embedded tissue blocks (FFPE) from tissue biopsies of 167 non-small cell lung carcinoma (NSCLC) patients and 167 healthy controls. The tissue biopsies were studied for mutations in exons 18-21 of the EGFR gene. This study was performed using PCR followed by DNA sequencing. We identified 63 mutations in 33 men and 30 women. Mutations were detected in exon 19 (delE746-A750, delE746-T751, delL747-E749, delL747-P753, delL747-T751) in 32 patients, exon 20 (S786I, T790M) in 16, and exon 21 (L858R) in 15. No mutations were observed in exon 18. The 63 patients with EFGR mutations were considered for upfront therapy with oral tyrosine kinase inhibitor (TKI) drugs and have responded well to therapy over the last 15 months. The control patients had no mutations in any of the exons studied. The advent of EGFR TKI therapy has provided a powerful new treatment modality for patients diagnosed with NSCLC. The study emphasizes the frequency of EGFR mutations in NSCLC patients and its role as an important predictive marker for response to oral TKI in the south Indian population.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6949-6954
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• 2014

Background: Recent studies have indicated that microRNA-15a (miR-15a) is dysregulated in breast cancer (BC). We aimed to evaluate the expression of miR-15a in BC tissues and corresponding para-carcinoma tissues. We also focused on effects of miR-15a on cellular behavior of MDA-MB-231 and expression of its target gene synuclein-${\gamma}$ (SNCG). Materials and Methods: The expression levels of miR-15a were analysed in BC formalin fixed paraffin embedded (FFPE) tissues by microarray and quantitative real-time PCR. CCK-8 assays, cell cycle and apoptosis assays were used to explore the potential functions of miR-15a in MDA-MB-231 human BC cells. A luciferase reporter assay confirmed direct targets. Results: Downregulation of miR-15a was detected in most primary BCs. Ectopic expression of miR-15a promoted proliferation and suppressed apoptosis in vivo. Further studies indicated that miR-15a may directly interact with the 3'-untranslated region (3'-UTR) of SNCG mRNA, downregulating its mRNA and protein expression levels. SNCG expression was negatively correlated with miR-15a expression. Conclusions: MiR-15a has a critical role in mediating cell cycle arrest and promoting cell apoptosis of BC, probably by directly targeting SNCG. Thus, it may be involved in development and progression of BC.

• 대한임상검사과학회지
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• 제50권2호
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• pp.197-204
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• 2018

다발성 골수종(MM)은 혈액 종양의 주요 사망 원인이다. 최근에 다발성 골수종 진단에 microRNA를 이용한 실험이 보고되고 있다. 이에 우리는 다발성 골수종 진단 마커로 miR-221을 이용 할 수 있는지 확인하고자 하였다. 본 연구는 다른 혈액학적 질환이 없는 다발성 골수종 환자 20명을 대상으로 하였다. MicroRNA 추출은 다발성 골수종 환자의 파라핀 포매 조직을 이용하였다. 우리는 다발성 골수종의 microRNA 표적 유전자로 miR-15a, miR-16, miR-21, miR-181a 그리고 miR-221을 선택하였다. 유의성 검정은 fold change 값을 기준으로 1.5이상 또는 -1.5 미만의 결과를 유의성이 있는 경우로 하였다. Fold change 값은 인간 유전자 SNORD43에 의해 표준화된 데이터를 기준으로 하였다. Fold change 값이 1.5 이상은 "overexpression", -1.5 미만의 값은 "underexpression"으로 정의되었다. miR-221의 65.0% (13/20)에서 "overexpression"으로 유의성이 있음을 확인하였고, 다발성 골수종 환자에서 형질세포가 30% 이상인 그룹과 이하의 그룹은 유의성을 보이지 않았다. MiR-221은 서구인과 같은 결과를 얻었으며, 다발성 골수종 환자에서 miR-221이 다발성 골수종 환자 진단에 매우 중요한 지표가 될 수 있을 것으로 생각된다. 결론적으로 miR-221은 한국인의 다발성 골수종 진단 또는 예후 지표로 활용할 수 있음을 확인하였다.

• 대한임상검사과학회지
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• 제52권4호
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• pp.381-388
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• 2020

정확한 분자병리검사를 위해 검체에 대한 질관리의 중요성이 강조되고 있다. 본 연구에서 BRAF, EGFR, KRAS 돌연변이 검사에 대해, 검체 종류 및 처리과정 중 DNA 품질에 영향을 미치는 인자를 알아보고자 하였다. 모두 1,772건의 분자병리검사에 대해, 검체의 종류, 포르말린 고정시간, 재검기록 등 DNA 품질에 영향을 미칠 수 있는 임상병리학적 요소를 조사하였다. 세포검체는 고정액으로 처리를 한 검체보다 생리식염수에 보관한 검체가 DNA 품질이 좋았다. 조직검체는 10% 중성 포르말린에 24시간 이내에 고정된 검체가 그 이상 고정된 검체보다 DNA 품질이 좋았다. 검체 종류 중 신선조직검체, 그리고 종양세포밀도가 높은 조직검체가 파라핀포매조직 및 세침흡인액상 세포검체보다 상대적으로 DNA 품질이 좋았다. 10% 중성 포르말린에 24시간 이상 오래 고정된 검체일수록 Non-PNA Ct값이 비례 증가하여, 포르말린 고정시간이 검체 DNA 품질에 영향을 준다는 것을 확인하였다. 결론적으로 종양세포의 밀도 및 적절한 포르말린 고정시간이 DNA 품질을 유지하는데 가장 중요한 요소이며, 신속하고 정확한 분자병리진단을 위해 이들 요소를 적절히 관리하여 최적의 DNA 품질을 유지하도록 해야 한다.