Pulmonary Epstein-Barr virus-associated smooth muscle tumor after kidney transplantation: two case reports with review of differential diagnosis.

Pulmonary nodules are a common complication in solid organ transplant recipients, and may have various underlying causes, with Epstein-Barr virus-associated smooth muscle tumor (EBV-SMT) being one of them. Given the rarity of this entity, we describe the diagnosis and therapeutic interventions for post-transplant EBV-SMT in two individuals. Both cases involved female patients who were diagnosed with multiple pulmonary nodules 60 months and 116 months, respectively, after receiving living-related kidney transplantation. Pathological examination revealed a spindle cell tumor, with immunophenotype and EBV in situ hybridization supporting the diagnosis of EBV-SMT. After diagnosis, these two patients underwent intervention by decreasing their intake of immunosuppressants. As of the latest follow-up, the patients' lesion size remained stable, and their overall condition was favorable. We also reviewed literature about the morphological and molecular pathological features of EBV-SMT and highlighted the diagnosis and differential diagnosis of pulmonary spindle cell lesions especially in the setting of immunosuppression.

Microvascular invasion is associated with poor survival in patients with dual-phenotype hepatocellular carcinoma.

OBJECTIVES: Microvascular invasion (MVI) has previously been reported to be related to cancer prognosis; however, its significance in patients with dual-phenotype hepatocellular carcinoma (DPHCC) remains uncharacterized. We studied the role of MVI in the survival of patients diagnosed with DPHCC in Fujian, China, which has a high incidence of HCC. METHODS: Patients with DPHCC (n = 84) who had undergone surgical interventions at the 900th Hospital of the Joint Logistic Support Force between 2013 and 2019 were retrospectively analyzed using the log-rank test and Kaplan-Meier method. Univariate and multivariate Cox model analyses were also conducted to further understand the correlation between MVI and patient survival. RESULTS: Our results indicated that MVI was related to poor survival. According to the univariate analysis, MVI, the number of tumor lesions, necrosis, differentiation, peripheral hepatic fibrosis, the expression of cytokeratin 19 (CK19), and serum levels of both ɑ-fetoprotein (AFP) and cancer antigen-199 showed a strong correlation with overall survival. Necrosis and serum AFP levels were strongly related to an increased risk of death, according to the multivariate analysis. Tumor size; the number of tumor lesions; differentiation; peripheral hepatic fibrosis; liver capsule invasion; and expression of CK19, vascular endothelial growth factor, CK7, and mucin 1 showed a correlation with MVI, per the outcomes of χ2 tests. CONCLUSIONS: Microvascular invasion may correlate with the survival of patients with DPHCC and could potentially serve as a prognostic predictor of survival.

LncRNA AFAP1-AS1 Modulates the Proliferation and Invasion of Gastric Cancer Cells by Regulating AFAP1 via miR-205-5p.

PURPOSE: The present study investigated the expression and function of the long noncoding RNA (lncRNA) actin filament associated protein 1 antisense RNA1 (AFAP1-AS1) related to gastric cancer (GC), based on previous results from a microarray analysis. METHODS: Real-time quantitative polymerase chain reaction (qPCR) was used to verify the expression of AFAP1-AS1 in 97 fresh GC tissues and paired non-GC tissues, as well as in six different GC cell lines (BGC-823, SGC-7901, MGC-803, AGS, MKN-45, and MKN-28). The expression levels were subsequently correlated with the clinicopathological features of patients. siRNA against AFAP1-AS1 was transfected into GC cell lines, and cell proliferation, migration, and invasion were detected before and after silencing of AFAP1-AS1 expression. Luciferase reporter gene analysis was used to confirm the target gene of microRNA-205-5p (miR-205-5p) in 293T cells. The potential mechanism was subsequently investigated. RESULTS: qPCR results showed that AFAP1-AS1 was significantly overexpressed in GC tumor tissues and also GC cell lines, comparing to their paired non-GC tissues. Furthermore, statistical analysis revealed that the overexpression of AFAP1-AS1 was significantly correlated with tumor size (p=0.018) and grade of differentiation (p=0.042). Subsequently, artificially decreasing the expression of AFAP1-AS1 with its specific siRNA dramatically inhibited the proliferation, migration and invasion of GC cell lines (SGC-7901 and BGC-823 cells). Mechanical analysis suggested that AFAP1-AS1 is involved in regulation of its maternal gene, AFAP1, at both mRNA level and protein level. Luciferase reporter gene assay indicated that lncRNA AFAP1-AS1, as a ceRNA, is able to sponge miR-205-5p. Moreover, miR-205-5p has been well demonstrated to participate in the regulation of AFAP1 expression and the phenotypes of GC cells, including proliferation, migration and invasion. CONCLUSION: AFAP1-AS1, as a novel biomarker of GC, promotes the proliferation migration and invasion of GC cells and function as ceRNA to target AFAP1 by sponging miR-205-5p.

LncRNA Expression Profiling in Advanced Resected Gastric Adenocarcinoma Tissues.

BACKGROUND: Long noncoding RNAs (lncRNAs) are the chief products of human transcriptomes and have a major function in mediating gene expression. Abnormal lncRNA levels have been detected in gastric cancer. However, changes in lncRNA expression in advanced gastric adenocarcinoma (GA) are largely unexplored. METHODS: We studied the expression of lncRNAs and mRNAs in 6 advanced resected GA (ARGA) tissues using a lncRNA microarray chip. RESULTS: Among 22,870 lncRNAs expressed in ARGA and paired nonneoplastic tissues (non-GA), 1,769 and 1,710 were up- or downregulated, respectively, in all 6 ARGA tissues (≥ 2.0-fold, p < 0.05). The expression of 5 differentially expressed lncRNAs, HNF1A-AS1, RP11-62F24.2, GAS5, MALAT1, and H19 were randomly selected to be measured in 47 patients using real-time quantitative reverse transcription PCR (qRT-PCR), and the data were consistent with those obtained from the microarray chip. Analysis of their nearby coding genes (mRNAs) revealed that the main associated GO (gene ontology) classes were genes that regulate cellular metabolic processes, protein binding and receptor binding, whereas the main associated pathways were MAPK signaling, which regulates cell proliferation and differentiation and the apoptosis pathway, which is cancer-related. Some (n = 37) differentially expressed lncRNAs had direct annotated functions; among these lncRNAs, 27 were associated with cancer, cancer pathways, oncogenes, and tumor suppressor genes and with cell development and differentiation. CONCLUSIONS: Expression differences in lncRNAs exist between advanced GA and noncancerous gastric tissues, so lncRNA expression patterns may explain gastric carcinogenesis and progression as well as serve as candidate biomarkers for the treatment of GA.

Long non­coding RNA RP1­163G9.1 is downregulated in gastric adenocarcinoma and is associated with a poor prognosis.

The aim of the present study was to investigate the expression, function and underlying molecular mechanism of the long non­coding (lnc) RNA RP1­163G9.1 in patients with gastric adenocarcinoma (GA). The expression levels of lncRNA RP1­163G9.1 were determined in 112 paired clinical GA tissues by reverse transcription­quantitative polymerase chain reaction analysis. Subsequently, the potential clinical values of lncRNA RP1­163G9.1 were analyzed with statistical methods. Additionally, the function of lncRNA RP1­163G9.1 was explored at the cellular level using the Cell Counting Kit­8 proliferation assay, Transwell experiments, fluorescence in situ hybridization (FISH), colony formation assay and flow cytometry. Furthermore, the function of lncRNA RP1­163G9.1 was assessed in vivo using subcutaneous tumorigenesis experiments in nude mice. lncRNA RP1­163G9.1 expression in GA tissues and cells was significantly decreased when compared with that in control gastric tissues (P<0.001) or gastric epithelial cells GES­1 (P<0.05). This finding was associated with the depth of invasion (P=0.001), lymph node metastasis (P=0.009), tumor size (P=0.037) and immunocytochemistry marker Ki­67 (P=0.010). FISH detection demonstrated that lncRNA RP1­163G9.1 was primarily located in the cytoplasm. Notably, overexpression of lncRNA RP1­163G9.1 significantly decreased cell proliferation (P<0.01), colony formation (P<0.01), invasion (P<0.01) and the number of cells at the S­phase of the cell cycle (P<0.05); However, it did not exert a significant effect on apoptosis (P>0.05). Furthermore, tumor formation experiments revealed that overexpression of lncRNA RP1­163G9.1 inhibited cancer cell proliferation in nude mice. The present research indicated that low expression of lncRNA RP1­163G9.1 may be associated with enhanced tumor proliferation and invasion in GA.

Low Serum Albumin Correlates with Poor Survival in Gastric Cancer Patients.

BACKGROUND: Preoperative nutritional status as evidenced by serum albumin measurement is associated with cancer prognosis but the clinical significance of this for patients with gastric cancer (GC) is unclear. Therefore, we evaluated an association between preoperative serum albumin and GC patient survival in the Fujian area which has a higher incidence for GC in China. METHODS: GC patients (n = 309) who underwent surgical treatment at Fuzhou General Hospital between 2010 and 2013 were retrospectively assessed using a Kaplan-Meier method and log-rank test, as well as univariate and multivariate Cox model analyses, to confirm a correlation between patient survival and preoperative serum albumin. RESULTS: Data show that low serum albumin was associated with poorer survival. Preoperative serum CEA, CA199, and albumin and tumor size, T staging, and lymph node metastases (LNM) were significantly associated with overall survival according to univariate analysis. Lower serum albumin (HR: 2.018, 95% CI [1.204 - 3.381], p = 0.008) and advanced cancers with deeper invasion (T3 + T4 stages) and with lymph node metastases were significantly associated with increased death risk according to multivariate analysis. Preoperative serum total protein, patient age, tumor size, T staging, and LNM were correlated with serum albumin according to chi-squared analysis. CONCLUSIONS: Preoperative serum albumin may be related to GC patient survival and may hold promise as a prognostic predictor for such survival.

Overexpression of Pyruvate Kinase Type M2 (PKM2) Promotes Ovarian Cancer Cell Growth and Survival Via Regulation of Cell Cycle Progression Related with Upregulated CCND1 and Downregulated CDKN1A Expression.

BACKGROUND Many findings have shown that pyruvate kinase type M2 (PKM2) plays crucial roles in regulating the occurrence and development of various human cancers; however, its roles in ovarian cancer oncogenesis remain to be determined. MATERIAL AND METHODS The expression intensity of PKM2 in ovarian cancer tissues was examined by immunohistochemistry (IHC), and was then correlated to patient clinicopathologic characteristics. The roles of PKM2 in ovarian cancer cell proliferation, growth, and survival were examined by CCK-8, colony forming, and flow cytometry assays. The potentially involved molecular were then investigated by Western blot analysis. RESULTS IHC results showed that PKM2 was overexpressed in 100 of 114 (87.7%) serous ovarian cancer tissues as compared with 50 cases of non-cancerous ovarian tissues, and was associated with tumor size ≥7.5 cm and <7.5 cm (p<0.05). Overexpression of PKM2 in SKOV3 and HEY ovarian cancer cells by transfection with PKM2 lentivirus vector led to increased cell proliferation, growth, and survival, which may be related with PKM2 being able to increase cell cycle progress: G1 stage decreased, whereas S stage significantly increased. In contrast, all functions of SKOV3 and HEY cells described above were reversed by knocked down PKM2 expression using siRNA. Further data showed that overexpressed PKM2 led to increased CCND1 and decreased CDKN1A expression, whereas underexpressed PKM2 led to decreased CCND1 and increased CDKN1A expression in ovarian cancer cells. CONCLUSIONS PKM2 may play important roles in ovarian cancer development and may be a treatment target for this cancer.

Evaluation of the expression and clinical value of lncRNA AC010761.9 in human gastric adenocarcinoma.

BACKGROUND: The current study determined the expression and clinical value of lncRNA AC010761.9 in human gastric adenocarcinoma (GA). METHODS: Real-time quantitative reverse transcription (qRT)-PCR was used to detect the level of lncRNA expression in 145 GA tissues and three GA cell lines, and the correlation between its level and clinicopathologic characteristics and potential corresponding mRNA of TNF receptor-associated factor 4 gene (TRAF4) was then evaluated. RESULTS: Elevated lncRNA AC010761.9 was detected in all 6 GA tissues by previous lncRNA expression profile microarray assay. LncRNA AC010761.9 was over-expressed in 99 of 145 GA tissues (68.3%) with an elevated fold change of up to 35.14 compared to matched paracancerous tissues (p < 0.05), and was also over-expressed in the 3 GA cell lines (MGC803, BGC823, and SGC7901) compared to the normal gastric mucosal epithelial cell line (GES-1 cells; p < 0.05) by qRT-PCR. The elevated expression of this lncRNA was related to tumor size (p = 0.028), degree of differentiation (p = 0.047), and serum carbohydrate antigen (CA19-9) and carcinoembryonic antigen (CEA) concentrations (p = 0.026 and p = 0.037, respectively). Multivariate analysis further confirmed that the expression of lncRNA AC010761.9 was related to the degree of tumor differentiation (p = 0.015). Additionally, the expression of lncRNA AC010761.9 had a positive correlation with the mRNA expression of the potentially associated gene (TRAF4) in GA tissues (r = 0.385, p < 0.01). CONCLUSIONS: LncRNA AC010761.9 may be linked to GA progression and is a potential new biomarker for GA.

Circular RNAs expression profiles in human gastric cancer.

Circular RNAs (circRNAs) are implicated in a variety of cancers. However, the roles of circRNAs in gastric cancer (GC) remain largely unknown. In the current study, circRNAs expression profiles were screened in GC, using 5 pairs of GC and matched non-GC tissues with circRNA chip. Preliminary results were verified with quantitative PCR (qRT-PCR). Briefly, total of 713 circRNAs were differentially expressed in GC tissues vs. non-GC tissues (fold change ≥ 2.0, p < 0.05): 191 were upregulated, whereas 522 were downregulated in GC tissues. qRT-PCR analysis of randomly selected 7 circRNAs from the 713 circRNAs in 50 paired of GC vs. non-GC control tissues confirmed the microarray data. Gene ontology (GO) and KEGG pathway analyses showed that many circRNAs are implicated in carcinogenesis. Among differentially expressed circRNAs, hsa_circ_0076304, hsa_circ_0035431, and hsa_circ_0076305 had the highest magnitude of change. These results provided a preliminary landscape of circRNAs expression profile in GC.

The function of homeobox genes and lncRNAs in cancer.

Recently, the homeobox (HOX) gene family has been reported as a factor in tumorigenesis. In the human genome, the HOX gene family contains 4 clusters with 39 genes and multiple transcripts. Mutation or abnormal expression of genes is responsible for developmental disorders. In addition, changes in the levels and activation of certain HOX genes has been associated with the development of cancer. Long non-coding RNAs (lncRNAs) have also been identified to serve critical functions in cancer. Although a limited number of lncRNAs have been previously investigated, the list of functional lncRNA genes has recently grown. Two of the most important and well-studied lncRNAs and HOX transcript genes are HOX transcript antisense RNA (HOTAIR) and HOXA distal transcript antisense RNA (HOTTIP). The present study aimed to review not only the function of the HOTAIR and HOTTIP genes in certain forms of cancer, but also to review other HOX genes and protein functions in cancer, particularly HOX family genes associated with lncRNAs.

Evaluation of the Clinical Significance of Serum Carcinoembryonic Antigen in Patients with Resectable Gastric Adenocarcinoma.

BACKGROUND AND AIMS: Carcinoembryonic antigen (CEA) is the most commonly used tumor marker for gastrointestinal cancers but its value for resectable gastric adenocarcinoma (RGA) patients in areas of high GA incidence is uncertain. METHODS: We retrospectively studied 400 subjects with RGA from the Fujian Province in China, which has a high incidence of GA. Patients had surgery between January 2010 and December 2013. CEA was measured and correlated to pathology. RESULTS: High pretreatment serum CEA (>5 ng/mL) was associated with patient age (p = 0.000), tumor size (p = 0.008), and T and N stages (p = 0.002, p = 0.032, respectively), alpha fetoprotein (p = 0.014), and CA19-9 (p = 0.000). High CEA was significantly associated with poor overall survival. Overall survival in the whole group of patients was 63.8%, whereas it was only 42.9% in the high CEA group (p = 0.0001). Mean overall survival for high CEA patients was significantly shorter than patients with low CEA (36.5 ± 2.63 months vs. 47.4 ± 0.98 months, p = 0.000). Multivariate analysis confirmed that pretreatment serum CEA was an independent prognostic factor for increased death risk. Additionally, mean CEA in 45 high CEA patients was reduced after surgery. CONCLUSIONS: Pretreatment serum CEA may help to predict survival for patients with RGA in high GA incidence areas.

The Value of Preoperative Neutrophil to Lymphocyte Ratio in Indicating Lymph Node Metastasis in Patients with Resectable T2 Stage Gastric Adenocarcinoma.

BACKGROUND: Lymph node metastasis (LNM) is closely associated with poor prognosis in patients with resectable T2 stage gastric adenocarcinoma (RT2-GA). Preoperative blood neutrophil to lymphocyte ratio (NLR) has been identified to be a very valuable predictor for prognosis in patients with diverse cancers. The aim of this investigation was to assess the relationship between NLR and LNM in RT2-GA. METHODS: This retrospective study reviewed 230 patients who underwent surgery for removal of primary T2-GA from August 2002 to December 2013 in a single hospital. Preoperative routine blood test data were collected and the relationship between NLR and LNM in RT2-GA was evaluated by X2 test and multivariate logistic regression analysis. RESULTS: The median value of NLR was 2.18 among 230 patients. Based on the median NLR value, the patients were categorized into two groups: low NLR group (NLR ≤ 2.18) and high NLR group (NLR > 2.18). χ2 test results exhibited that the preoperative NLR was significantly associated with the numbers of metastatic lymph nodes (≤ 6 and > 6) (p = 0.003) and status of lymph node involvement (N0, N1, and N2 stage) (p = 0.032). Multivariate analyses further confirmed that NLR > 2.18 was significantly associated with increased risk of appearing more numbers of metastatic lymph node or higher N stage which exhibited a 4.15- or 7.09-fold elevated risk compared to that of NLR ≤ 2.18. CONCLUSIONS: The preoperative NLR is closely associated with LNM in patients with RT2-GA, which may be used as a predictor indicating more serious LNM in this type of cancer.

Elevated Preoperative Platelet to Lymphocyte Ratio Indicates Poor Survival in Patients with Resected High-grade Serous Ovarian Carcinoma.

BACKGROUND: Platelet to lymphocyte ratio (PLR) is widely used as an inflammation-related cancer biomarker. However, since its prognostic importance in resected high-grade serous ovarian carcinoma (HGSC) is still unknown, we investigated the association between PLR and the prognosis in resected HGSC in this study. METHODS: Details of 103 patients with HGSC who underwent ovarian resection were collected in this retrospective study. Preoperative PLR was calculated based on platelet and lymphocyte count values. A χ2 test was used to analyze the relationship between PLR and clinical variables, a Kaplan-Meier curve and log rank analysis was used to evaluate overall survival, and multivariable analysis was used to analyze the prognostic factors. RESULTS: The preoperative PLR median value (188.8) was used to divide patients into two groups: the high PLR group (PLR > 188.8) and low PLR group (PLR ≤ 188.8). A high PLR was significantly associated with a higher death rate (81.6% vs. 59.3%, p = 0.013) and a shorter median overall survival time (37 months vs. 58 months, p = 0.035) during follow-up (median length = 43 months). Multivariable data further demonstrated that a high PLR was related to a two-fold increase in risk of death (hazard ratio [HR]: 2.19, 95% confidence interval (CI): 1.30 - 3.68, p = 0.003). In addition, the risk of a CA125 of > 640.0 U/mL was significantly greater in the high PLR group (odds ratio [OR]: 2.72, 95% CI: 1.18 - 6.27, p = 0.019). Multivariable analysis suggests that PLR was an independent prognostic factor for resected HGSC. CONCLUSIONS: PLR has potential as a prognostic biomarker for predicting the survival of patients with resected HGSC.

Expression and clinical significance of long non-coding RNA HNF1A-AS1 in human gastric cancer.

BACKGROUND: Increasing evidence has demonstrated that long non-coding RNAs (lncRNAs) play essential roles in the occurrence and development of human cancers, including gastric cancer (GC). However, the functional and clinical significance of lncRNAs are still poorly understood. METHODS: In this study, the expression of LncRNA HNF1A antisense RNA 1 (HNF1A-AS1) was first examined by lncRNAs microarray analysis in 6 GC tissues, and was then further verified by real-time quantitative reverse transcription PCR (qRT-PCR) both in 3 GC cell lines and 161 cases of GC tissues. We also evaluated the association between HNF1A-AS1 expression and clinicopathological features of patients with GC. RESULTS: LncRNAs microarray analysis results exhibited that HNF1A-AS1 was downregulated in GCs tissues (mean fold change 2.06, p < 0.05), which was further confirmed by qRT-PCR. The results from qRT-PCR showed that the expression of HNF1A-AS1 was not only downregulated in three GC cell lines (AGS, BGC-823, and MKN-45) relative to that in a normal gastric mucosal epithelial cell line (GES-1), but also decreased in GC tissues relative to that in paired adjacent non-neoplastic tissues (low expression, 94 of 161; low expression rate, 58.38%). Furthermore, low HNF1A-AS1 expression was associated with tumor size/diameter (p = 0.005, multivariate analysis), levels of serum carcinoembryonic antigen (CEA), and carbohydrate antigen 19-9 (CA19-9), and RRM1 expression in tissue samples (p = 0.028, p = 0.009, and p = 0.006, respectively). CONCLUSIONS: Taken together, our data indicate that lncRNA HNF1A-AS1 may be a regulator of GC, and thus, it may have potential as a novel biomarker and treatment target for this type of cancer.

High Expression of IncRNA-PCNA-AS1 in Human Gastric Cancer and its Clinical Significances.

BACKGROUND: Long noncoding RNAs (IncRNAs) have been demonstrated to be associated with human cancer. However, the clinical value of most lncRNAs to serve as the biomarkers for gastric cancer remains largely unknown. In this study, we examined the relationship between the expression levels of IncRNA, PCNA antisense RNA 1 (PCNA-AS1), and clinicopathological characteristics of gastric cancer. METHODS: A total of 90 tissue samples from patients with gastric cancer were collected, and the IncRNA PCNA-AS1 levels in cancer and paired non-cancer tissues was detected by real-time quantitative RT-PCR (qRT-PCR). The relationship between PCNA-AS1 levels and the clinicopathological characteristics were then evaluated. RESULTS: PCNA-AS1 expression levels were increased in 68 of 90 gastric cancer tissues (75.56%) and were associated with invasion (p = 0.038), but not associated with other clinicopathological characteristics. The expression levels of PCNA-AS1 were also related with immunohistochemical biomarkers of BRCA1 (p = 0.043) and RRM1 (p = 0.023). CONCLUSIONS: These data indicated that IncRNA-PCNA-AS1 may participate in the gastric cancer carcinogenesis and development and may serve as a new biomarker for patients with gastric cancer.