Relationship between driver gene mutations, their relative protein expressions and survival in non-small cell lung carcinoma in Macao.

OBJECTIVES: We report the status of most common gene mutations in non-small cell lung carcinoma (NSCLC) in Macao, and explore the relationship between each gene mutation and clinicopathologic features and survival. METHODS: EGFR, KRAS and BRAF mutations were detected by PCR in 122 cases of NSCLC. ALK translocation and MET amplification were detected by fluorescence in situ hybridization (FISH). MET and thyroid transcription factor (TTF-1) were investigated by immunohistochemistry. Clinical data were collected for analyzing their correlation with the gene mutations. RESULTS: The mutation of EGFR, KRAS and BRAF was detected in 48 (39.3%), 13 (10.7%) and 3 (2.5%) of 122 cases of NSCLC, respectively. ALK translocation and MET amplification were detected in 7 (5.7%) and 3 cases (2.5%). The rate of EGFR mutation was significantly higher in female and non-smoker patients. In TTF-1 positive cases EGFR mutation was more frequent. Age of the patients over 62-year old was correlated with KRAS mutations. The concordance between ALK IHC and FISH was 58.3%. The MET protein in the cases with MET amplification was 100% positive. The survival was lower in the patients with positive MET protein than those with negative. MET protein was an independent prognostic factor for NSCLC. CONCLUSIONS: EGFR mutation occurred frequently in the female never smoke patients with NSCLC. KRAS mutation was more common in old patients. Negative MET protein expression could be used as a negative predictive marker of MET amplification. MET protein expression was an independent prognostic factor for NSCLC.

[Glypican 3 expression in hepatoblastoma and its diagnostic implication].

OBJECTIVE: To explore the expression and diagnostic significance of glypican-3 (GPC3) in hepatoblastoma. METHODS: Five tissue microarray paraffin blocks were constructed to include 54 cases of hepatoblastoma. The tumor tissue samples were obtained from 3 surgical biopsies, 33 needle biopsies, 5 stage I resection tumors, and 13 stage II resection tumors after transcatheter arterial chemoembolization. Ten samples of non-neoplastic hepatic tissue adjacent to tumor were used as control. Immunohistochemical staining of GPC3 (clone 1G12) was performed. Among the 54 cases of hepatoblastoma, 22 cases were fetal subtype, 24 cases were mixed fetal and embryonal subtype and 8 cases were mixed epithelial and mesenchymal type. RESULTS: GPC3 was positive in fetal epithelial cells (54/54, 100%), but negative or weakly positive in embryonic epithelial cells in all cases of hepatoblastoma. Undifferentiated small cells and all mesenchymal components were negative for the expression. Non-neoplastic hepatocytes adjacent to tumor were negative for GPC3 expression (0/10) . CONCLUSIONS: Fetal epithelial components of hepatoblastoma express GPC3 protein detectable by immunohistochemistry. Normal hepatocytes after birth, small cell undifferentiated and embryonic epithelial components of hepatoblastoma do not or weakly express GPC3 protein. Therefore, GPC3 immunohistochemistry offers a valuable aid to the diagnosis of hepatoblastoma in infants and children.

[Amplification of HER2 gene in gastric carcinoma detected by dual in-situ hybridization].

OBJECTIVE: To explore the status of HER2 gene amplification and its product HER2 protein expression in gastric carcinoma, so as to aid in patient selection for anti-HER2 targeted chemotherapy. METHODS: Eighty-five cases of gastric carcinoma biopsy tissues were collected. The status of HER2 gene amplification was detected by dual in situ hybridization (dual-ISH). And HER2 protein was detected by immunohistochemistry. RESULTS: HER2 gene amplification was detected in 10/85 (11.8%) cases of gastric carcinoma, and no amplification was detected in 75/85 (88.2%) cases. In the 10 cases with HER2 amplification, HER2 immunoreaction scorings of 3+, 2+ and 0/1+ were present in 7, 2 and 1 cases, respectively. In the 75 cases without HER2 amplification, HER2 immunoreaction scorings of 3+, 2+ and 0/1+ were present in 0, 18 (24.0%) and 57 (76.0%) cases, respectively. Histologically, most gastric carcinoma with amplification of HER2 gene was moderately differentiated tubular adenocarcinoma. CONCLUSIONS: HER2 gene dual-ISH technique is a reliable and objective method for detecting HER2 gene amplification in gastric carcinoma biopsy. Clinically, only few gastric carcinomas show HER2 gene amplification and are suitable candidates for anti-HER2 targeted chemotherapy.

[Significance of glypican-3 immunohistochemistry in diagnosis of hepatocellular carcinoma].

OBJECTIVE: To explore the diagnostic significance of glypican-3 (GPC3) immunohistochemistry in hepatocellular carcinoma (HCC). METHODS: Fourteen tissue microarray paraffin blocks were constructed, which comprised 731 samples from hepatic tumors and paratumor tissues, including 357 cases of HCC, 26 cholangiocarcinoma, 171 HCC adjacent hepatic tissue including cirrhosis, 93 hemangioma adjacent hepatic tissues, and 84 carcinomas metastatic to liver. GPC3 (Clone 1G12) protein was detected immunohistochemically in all of cases with positive controls. RESULTS: GPC3 protein was positive in 72.0% HCC (257/357), but negative in the rest 374 of non-HCC cases, including cholangiocarcinoma, HCC adjacent hepatic tissue including cirrhosis, hemangioma adjacent hepatic tissues and metastatic carcinomas. GPC3 positive percentage was significantly correlated with histological grading of HCC (P < 0.01), highest in grade 3 (77.1%, 64/83) followed by grade 2 (73.3%, 187/255), grade 1 (6/12) and grade 4 (0). CONCLUSIONS: GPC3 is a valuable diagnostic marker for hepatocellular carcinoma with sensitivity of 72.0%, and a differential diagnostic marker from tumor adjacent hepatic tissue and carcinomas metastatic to liver with specificity of 100%.

Prevalence and genotype distribution of cervical human papillomavirus infection in Macao.

Population-specific epidemiological data on human papillomavirus (HPV) infection are essential for formulating strategies to prevent cervical cancer. The age-specific prevalence of HPV infection was determined among 1,600 women enrolled for cervical screening in Macao. A U-shaped age-specific prevalence curve with a first peak (prevalence rate, 10%) at 20-25 years and a second peak (13%) at 51-55 years was observed. Co-infections with multiple types were detected in 32.5% of HPV-positive subjects and without significant variation among different age groups (P = 0.318). The majority (84.6%) of the positive samples harbored high- or probable high-risk HPV types, and these types also exhibited a similar U-shaped age-specific prevalence curve. In contrast, low and unknown-risk HPV types remained at a low prevalence (1.5-2.5%) throughout the age groups between 20 and 50 years, and with a small peak (4.5%) at 51-55 years. HPV 52 was the most common type found in 26.8% of positive samples, followed by HPV 16 (15.5%), HPV 68 (11.4%), HPV 18 and HPV 58 (8.9% each), HPV 54 (8.1%), HPV 53 (7.3%), HPV 39 (6.5%), HPV 33 and HPV 66 (5.7% each). In conclusion, because of the early peak of infection, vaccination and educational campaigns in Macao should start early and target at teenagers. The presence of a second peak containing mainly high-risk HPV types in older women indicates the need to evaluate the cover of the cervical screening programme for older women. Further study to determine the contribution of HPV 52 in high-grade cervical neoplasia and invasive cancers in Macao is warranted.

Distribution of human papillomavirus genotypes among cervical intraepithelial neoplasia and invasive cancers in Macao.

Macao is a densely populated city situated in East Asia where a relatively high prevalence of human papillomavirus (HPV) types 52 and 58 has been reported in women with invasive cervical cancer. To provide data for a population-specific estimation on the impact of HPV vaccines, paraffin-embedded tissues collected from women with invasive cervical cancer or cervical intrapeitheilal neoplasia grade 2 or 3 confirmed histologically were examined for HPV using the INNO-LiPa kit. Of the 35 HPV-positive patients with invasive cancer, one HPV type was detected in 68.6%, and 31.4% were co-infected with more than one HPV type. Overall, HPV 16, HPV 18, HPV 52, and HPV 54 were the most common types found respectively in 57.1%, 17%, 11.4%, and 8.5% of patients with invasive cervical cancer. Among the 59 HPV-positive patients with cervical intraepithelial neoplasia grade 2/3, 55.9% hardbored one HPV type, and 44.1% had co-infections. The common HPV types found included HPV 16 (52.5%), HPV 52 (23.7%), HPV 58 (18.7%), and HPV 33 (17%). Although HPV 11 (a low-risk type) was also found commonly in invasive cervical cancers (14.3%) and cervical intraepithelial neoplasia grade 2/3 (15.3%), the fact that they all existed as co-infections with another high-risk type suggested HPV 11 was not the cause of the lesion. The current vaccines targeting HPV 16/18 are expected to cover 62.9-74.3% of invasive cervical cancers and 32.2-55.9% of cervical intraepithelial neoplasia 2/3 in Macao. Widespread HPV vaccination is expected to reduce substantially the disease burden associated with cervical neoplasia in Macao.

Diagnostic utility of glypican-3 for hepatocellular carcinoma on liver needle biopsy.

AIMS: Glypican 3 (GPC3) is a heparan sulphate proteoglycan that shows elevated levels in the serum of patients with hepatocellular carcinoma (HCC), but not in healthy blood donors or patients with benign liver disease. This study explores the value of GPC3 expression for diagnosis of HCC by immunohistochemistry in liver needle biopsy specimens. METHODS: Archival material of liver needle biopsies from 54 patients with HCC, nine with focal nodular hyperplasia or focal liver cell dysplasia, five with cirrhosis, seven with hepatitis B or unremarkable liver tissue, seven with cholangiocarcinoma, and 30 with metastatic tumours, was retrieved for immunohistochemical staining with GPC3 antibody and appropriate positive and negative controls. RESULTS: Forty-five out of 54 cases of HCC showed positive GPC3 staining (83.4%). In contrast, all 58 non-HCC cases of liver biopsies, including focal nodular hyperplasia, focal liver cell dysplasia, cirrhosis, hepatitis B or unremarkable liver tissue, cholangiocarcinoma and metastatic tumours, were negative for GPC3. The sensitivity and specificity of GPC3 in HCCs were 83.4% and 100%, respectively. CONCLUSIONS: GPC3 is a valuable diagnostic marker for diagnosing HCC on liver needle biopsy. It can be used to distinguish HCC from other benign hepatic conditions and metastatic tumours in the liver.