1.Expression of c-MET, EGFR and HER-2 of human gastric adenocarcinoma
Lin LI ; Yao FU ; Hong-Yan WU ; Jun CHEN ; Jie-Yu CHEN ; Qi SUN ; Xin-Yun XU ; Yi-Hua WANG ; Xiang-Shan FAN
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):6-10
Purpose To investigate the expression status and their clinical significances of c-MET, epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor-2 (HER-2) in gastric adenocarcinoma (GC). Methods Tissue samples from 442 cases of GC in patients who accepted D2/D3 radical gastrectomy with R0 resection were stained by immunohistochemistry against c-MET, EGFR and HER-2.Results Over expression of c-MET, EGFR and HER-2 was identified in 195/442 (44.1%), 47/442 (10.6%) and 152/442 (34.4%) GC patients, respectively. Over expression of cMET was more often identified in GC patients with deeper T (P= 0.016), nerve involvement (P = 0.006) and the Lauren diffuse type (P = 0.029). EGFR in cases with vessel permeation (P = 0.012), and HER-2 in cases of distant metastasis (P =0.031), non-nerve involvement (P = 0.024), the Lauren intestinal type (P < 0.001) and G1/G2 grade (P < 0.001). Conclusion The receptors tyrosine kinase (RTKs) markers of cMET, EGFR and HER-2 might involve in the advance of GC, such as invasion and metastasis. The expression status of them could be used as the risk prediction and even the basis for future personalized therapy, especially for tyrosine kinase inhibitor (TKI) therapy, for GC patients.
2.Expression and clinical significance of N-Myc and p53 in prostate cancer tissues
Gui-Fang HE ; Xiao-Juan HUANG ; Lu-Lu ZHENG ; Li ZHANG ; Ling-Fan XU ; Chao-Zhao LIANG ; Yong-Ping CAI ; Yu YIN
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):10-13
Purpose To detect the expression of N-Myc and p53 in the tissues of prostate cancer (PCa) patients and to explore the relationship between them and their significance.Methods A total of 63 patients with PCa and 50 patients with benign prostatic hyperplasia (BPH) who underwent prostate surgery at the First Affiliated Hospital of Anhui Medical University were recruited in 2015-2016. The expression of N-Myc and p53 in pathological tissues were detected by immunohistochemistry of MaxVision method. Results The expression of N-Myc and p53 in PCa tissues was increased (P < 0.05). The expression of N-Myc and p53 in PCa tissues was correlated with bone metastases and TNM stage (P < 0.05), but not related to patient age, preoperative PSA level and other factors (P> 0.05). In addition, the expression of p53 was also correlated with Gleason score.Conclusion The high expression of N-Myc and p53 in PCa may involved in the malignant progression and metastasis of prostate cancer, and it is expected to become a new target for detecting PCa metastasis.
3.Application of D2-40/CD34-CK cocktail antibodies for colorectal cancer with insufficient lymph node harvest
Xian-Hua LIU ; Ying-Hao YU ; Xing-Feng QI ; Zai-Zeng WU ; Shun-Qi HU ; Xi-Sheng XIONG ; Juan XIANG ; Zhi-Yong ZHENG ; Li-Juan QU ; Xian-Zong YE
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):14-18
Purpose To investigate the value of application of D2-40/CD34-CK cocktail antibodies by double immunohistochemical staining for assessment of lymphovascular invasion (LVI) and to determine its prognostic significance in colorectal cancer with insufficient lymph node harvest. Methods Specimens from 133 cases of colorectal cancer with less than 12 lymph nodes were selected. HE staining and double immunohistochemical staining of the cocktail antibodies were performed to compare the difference of the two methods in screening for LVI. The The relationship between LVI confirmed by cocktail antibody immunohistochemical staining and clinicopathological characteristics and overall survival (OS) of patients was analyzed. Results (1) The detection rates of cocktail antibody double immunohistochemical staining and HE staining for LVI were 42.9% (57/133) and 21.8% (29/133) with statistically significant difference (P < 0.001). (2) The presence of LVI confirmed by double staining was significantly associated with Dukes staging, depth of invasion, clinical stages, lymph node metastasis and tumor budding (P < 0.05). (3) The presence of LVI, the location and extent of LVI, and the number of tumor cells in thrombus ≥5.5 for cases with LVI ≤2 clusters, were significantly associated with OS (P < 0.05). Conclusion D2-40/CD34-CK cocktail antibodies double staining is superior to routine HE staining in assessing LVI. LVI is intimately associated with tumor stage, lymph nodes metastasis and tumor budding, and it is an independent prognostic factor for CRC patients. It should be a supplementary examination for these patients with insufficient lymph node harvest.
4.Effects of adenosine on brain metastasis of lung cancer and its possible mechanism
Ming-Ke TANG ; Dian-Fang WEI ; Yang LIU ; Chao-Yue ZHANG ; Li-Juan QIN
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):19-22
Purpose To clarify the effect of adenosine on brain metastasis of lung cancer and the possible mechanism of adenosine promoting brain metastasis of lung cancer. Methods Western blot was used to dynamically detect the expression level of hypoxia inducible factor-1 (HIF-1) in lung cancer cells and tight junction protein ZO-1 in brain microvascular endothelial cells on blood-brain barrier. The content of adenosine in lung cancer cell culture was determined by ELISA. Fluorescence analysis was used to detect the changes of permeability of the blood-brain barrier model in vitro. Hemocytometer counts the number of A549 lung cancer cells in Transwell's lower chamber. Results The expression level of HIF-1 in lung cancer cells and the content of adenosine in lung cancer cell culture reached the highest level when lung cancer cells were deprived of oxygen for 12 hours. At the same time, the expression level of ZO-1 protein in the blood-brain barrier was the lowest, the blood-brain barrier permeability was the highest (7.11), and the number of lung cancer cells passing through the blood-brain barrier model was the highest (84.6). The permeability of the blood-brain barrier model increased after the action of adenosine, and its change trend was consistent with the effect of hypoxic lung cancer cell culture solution. Conclusion Hypoxia can induce the lung cancer cell to release adenosine, the increased adenosine can reduce the expression of tight junction protein ZO-1 in blood brain barrier, which leads to the increase of permeability of blood-brain barrier and eventually lead to brain metastasis of lung cancer.
5.Expression of MYB protein in adenoid cystic carcinoma of the salivary gland
Yu WANG ; Jun-Jie TANG ; Hong-Wei YUAN ; Yong-Feng JIA ; Lin SHI
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):23-27
Purpose To investigate the role of MYB in adenoid cystic carcinoma (ACC) of different origins and grades.Methods A total of 103 specimens of salivary gland tumors with complete pathologic data were collected from the Department of Pathology, Affiliated Hospital of Inner Mongolia Medical University. 64 cases of ACC of the salivary gland, which included 19 cases of grade I, 27 cases of grade Ⅱ, and 18 cases of grade Ⅲ were selected as experimental group. 7 cases of exrras-alivary ACC and 32 cases of non-ACC benign and malignant tumor of salivary gland were served as control group. The MYB protein expression was detected by MYB immunohistochemical En Vision two-step staining for both experimental and control group. Results The positive expression rates of MYB in 64 cases of salivary ACC and 7 cases of extrasalivary ACC were 54.69% (35/64) and 57.14% (4/7), respectively, with no statistically significant difference (P> 0.05). The positive expression rates of MYB in 64 cases of salivary ACC and 32 cases of non-ACC salivary benign and malignant tumors were 54.69% (35/64) and 6.25% (2/32), respectively, which showed statistically significant difference (P < 0.05). The positive expression rates of MYB were 68.42% (13/19), 66.67% (18/27) and 14.29% (4/18) in 64 cases of salivary ACC Ⅰ, Ⅱ and Ⅲ, respectively, and the difference was significant (P <0.05). The difference between grade Ⅰ and Ⅱ was not statistically significant (P> 0.016 7), while compared with grade Ⅲ, there was a statistical increase of positive expression rate of MYB in both grade Ⅰ and Ⅱ (P < 0.016 7). Conclusion MYB is highly expressed in grade I and Ⅱ of the salivary ACC and low expressed in grade Ⅲ. It is speculated that there may be other fusion gene mutation in salivary ACC grade Ⅲ. MYB is lowly expressed in non-ACC benign and malignant tumors of salivary gland, suggesting that MYB protein expression has a certain diagnostic value for the diagnosis of salivary ACC.
6.Expression of Fascin-1 and β-catenin protein in colorectal adenocarcinoma and its relevance with K-ras gene mutations
Gui MA ; Lan WANG ; Shu-Ping MA ; Fang BIAN ; Yan-Ni REN ; Qing-Rong HU ; Rong YANG
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):27-32
Purpose To examine the expression of Fascin-1 and β-catenin protein and K-ras gene mutation in colorectal adenocarcinoma, and to explore their role in progression of colorectal neoplasm and their relevance. Methods Fascin-1 and β-catenin were analyzed by use of immunohistochemistry En Vision two-step. K-ras gene mutation was detected by ARMS method.Relationship between overexpression of Fascin-1, the nuclear expression of β-catenin, and the mutations of K-ras gene and clinicopathologic parameters was analyzed, the correlation between them was also analyzed. Results In 112 colorectal adenocarcinoma samples, the overexpression rate of Fascin-1 protein was 27.7% (31/112), significantly higher than non-neoplastic mucosa (P < 0.01). The high nuclear expression rate of β-catenin was 29.5% (33/112) in adenocarcinoma and non-neoplastic mucosa respectively with a significant difference between two groups (P < 0.01). High expression rate of Fascin-1 protein and β-catenin were correlated significantly with lymph node metastasis (P = 0.022, P = 0.027), and TNM staging (P =0.042, P = 0.019) in colorectal adenocarcinoma. The overexpression of Fascin-1 protein was correlated with tumor location (P = 0.004). The mutation rate of K-ras gene was 34.8% (39/112), which showed no correlation with age, gender, tumor size, grade of differentiation, lymph node metastasis and TNM staging (P> 0.05). There was a correlation between the overexpressison of Fascin-1 protein, the nuclear expression of β-catenin and the mutation of K-ras gene (rs= 0.252, rs= 0.258, P < 0.05). The overexpression of Fascin-1 protein positively correlated with the nuclear expression of β-catenin (rs= 0.213, P < 0.05). Conclusion Fascin-1 protein and β-catenin protein are involved in invasion and metastasis of colorectal cancer and are associated with K-ras gene mutation. K-ras may promote the overexpression of Fascin-1 by virtue of nuclear expression ofβ-catenin, which provided a new research direction on the treatment of K-ras gene mutated colorectal adenocarcinoma.
7.Effect of up-regulation of miR-92a on the proliferation and angiogenesis of non-small cell lung cancer
Jian-Hua TANG ; Ping FENG ; Zhi-Hua ZHANG ; Bu WANG
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):38-42
Purpose To investigate the effect of down-regulation of miR-92 a on the proliferation and angiogenesis of nonsmall cell lung cancer (NSCLC). Methods Human NSCLC cell A549 was divided into three groups: A549 group (non-transfected A549 cells), sc-siRNA group (A549 cells transfected with sc-siRNA) and miR-92a-siRNA group (A 5 4 9 cells trans-fected with miR-92a-siRNA). The relative expression level of miR-92 a, PTEN and vascular endothelial growth factor (VEGF) in A549 cells and human bronchial epithelial (HBE) cells were detected by RT-PCR and Western blot respectively. The proliferation ability of A549 cells in each group was detected by living cell count and crystal violet staining experiment. Results The relative expression of miR-92 a in A549 cells was significantly higher than that in HBE cells (P < 0.05), the expression level of PTEN protein in A549 cells was significantly lower than that in HBE cells (P < 0.05), and the expression level of VEGF protein was significantly higher than that in HBE cells (P < 0.05).In the miR-92a-siRNA group, the relative expression of miR-92 a decreased (P < 0.05), the expression level of PTEN protein in-creased (P < 0.05), and the expression level of VEGF protein decreased (P < 0.05). The expression levels of PI3 K and Akt in miR-92a-siRNA group decreased (P < 0.05). the number of cells and cell proliferation ability in miR-92a-siRNA group reduced. Conclusion The expression of miR-92 a in NSCLC A549 cells is up-regulated, miR-92 a gene silencing can significantly inhibit cell proliferation and inhibit cell angiogenesis, PTEN and VEGF related PI3K/Akt signaling pathways may play an important role in this process.
8.Expression of long-chain non-coding RNA-LINC00485 in lung cancer and its effect on proliferation and migration of lung cancer cells
Xiang-Hui WANG ; Jiang-Ping HUANG ; Feng-He CUI ; Hai-Yun QIAN
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):42-46
Purpose To observe the expression of longchain non-coding RNA-LINC00485 (LINC00485) in lung cancer cell lines and tissues, and to investigate its effect on the proliferation and migration of lung cancer cells and its mechanism.Methods Quantitative real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect differential expression of LINC00485 in four lung cancer cell lines (H1975, A549, HCC827, H1299), normal alveolar epithelial cells HPAEPIC, and in 12 cases lung cancer tissues and adjacent tissues. Bioinformatics methods were used to predict the microRNA (miRNA) that LINC00485 may bind and target gene that miRNA may bind. Small interfering RNAs (siRNAs) that target silencing LINC00485 were transfected into HCC827 cells by liposomes.The expression levels of LINK00485, miR-361-5p, and p21 activated protein kinase 2 (PAK2) mRNA were detected by qRTPCR. The expression level of PAK2 protein was detected by Western blot. The cell proliferation ability was measured by MTS assay. Cell scratch assay was used to detect cell migration. Results Compared with normal alveolar epithelium, LINC00485 was highly expressed in lung cancer cell lines (P < 0.05), and the expression level was highest in HCC827 cells. The expression of LINC00485 in lung cancer tissues was higher than that in adjacent tissues (P < 0.01). After down-regulation of LINC00485 expression in HCC827 cells, the expression of miR-361-5p was up-regulated (P < 0.01), the expression of PAK2 mRNA and protein was down-regulated (P < 0.01), the proliferative capacity of HCC827 cells was decreased (P < 0.05), and the ability of cell migration was decreased (P < 0.01).Conclusion The expression of LINC00485 is increased in lung cancer cell lines and tissues. Down-regulation of LINC00485 can inhibit the proliferation and migration of lung cancer HCC827 cells by regulating the expression of miR-361-5p and PAK2 genes.
9.Comparison of immunohistochemical stain and 14C urea breath test in the diagnosis of helicobacter pylori associated gastritis
Wei-Song WAN ; Le WANG ; Yu-Chang HU ; Yu-Fei LIU ; Dong-Ping ZHENG ; Qing-Long HU
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):47-50
Purpose To evaluate the value of immunohistochemical stain (IHC) in the diagnosis of helicobacter pylori (HP) -associated gastritis with gastric biopsy tissue. Method239 cases of gastric biopsy were selected. All patients had14 C urea breath test (UBT) at the time of gastric biopsy. IHC for HP was performed with the biopsy tissues. The results were compared with gold standard results. Gold standard was established according to the Chinese Fourth National Consensus report on HP infection with modification. It was considered positive if two or more of the three test results were positive, including IHC, UBT, and HE. Results 239 gastric biopsy specimens fit the criteria. Compared with the gold standard, the sensitivity and specificity value of IHC stains were 97.3% and 100.0% respectively, higher than 91.1%, 81.9% of 14 C UBT, and 80.4%, 92.9% of HE stain respectively. The14 C UBT showed 9.6% false positive rate, and 4.2% false negative rate. Conclusion IHC is a valuable test in the diagnosis of HP-associated gastritis, and it should be recommended for routine histopathologic diagnostic workup if HP gastritis is clinically suspected or chronic active gastritis is identified in the biopsy tissue.
10.Expression of CBX8 in gastric adenocarcinoma and the relationship with clinicopathologic characters and survival prognosis
Xue-Na XIE ; Xuan ZHOU ; Yun-Qing CHEN ; Hong LI ; Feng-Gang XIANG
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):51-55
Purpose To explore the expression of CBX8 in gastric adenocarcinoma tissue and its clinical significance.Methods 119 cases of primary gastric adenocarcinoma treated by surgery and corresponding adjacent normal tissues were collected. The protein expression levels of CBX8 in gastric adenocarcinoma and their corresponding adjacent normal tissue was determined using tissue microarray and immunohistochemistry PV 6000 method staining. The relationship between CBX8 expression and clinicopathologic characters and survival prognosis was analyzed. Results The positive expression rate of CBX8 in gastric cancer and their corresponding adjacent normal tissue was 46.2% (55/119) and 13.4% (16/119) respectively. There was statistically significant difference between two groups (χ2=30.53, P < 0.01). The expression of CBX8 in gastric adenocarcinoma tissue was obviously correlated with the differentiation, clinical staging, and lymph node metastasis (P < 0.05). The high expression rate of CBX8 in the poorly differentiated gastric adenocarcinoma was lower than moderately differentiated and well differentiated group. The high expression rate of CBX8 in stage Ⅰ + Ⅱ gastric adenocarcinoma was higher than that in stage Ⅲ + Ⅳ. The gastric adenocarcinoma patients with high expression of CBX8 had low metastasis rate. Cox multivariate analysis showed CBX8, clinical staging, and lymphatic metastasis were independent predictors of overall survival and disease-free survival (P < 0.05). Kaplan-Meier analysis showed that the patients with higher expression of CBX8 had both longer overall survival time (P = 0.004) and disease-free survival time (P =0.004). Conclusion The expression of CBX8 may be involved in the tumorigenesis and progression of gastric adenocarcinoma. CBX8 is one of the independent predictor of prognosis, and the detecting of CBX8 expression may have important clinical value for the evaluation of gastric adenocarcinoma. CBX8 may became a new target for target therapy of gastric adenocarcinoma.