Effects of Minimally Invasive Esophagectomy and Open Esophagectomy on Circulating Tumor Cell Level in Elderly Patients with Esophageal Cancer.

OBJECTIVE: To investigate the effects of minimally invasive esophagectomy (MIE) and open esophagectomy (OE) on circulating tumor cell (CTC) level of elderly patients with esophageal cancer (EC). METHODS: A total of 78 elderly EC patients who aged over 64 years were divided into the MIE group (n = 40) and the OE group (n = 38). CTC enrichment was performed through CD326 (EpCAM) immunomagnetic beads positive sorting, and then labeled by CK-PE and CD45. The quantity of CTCs was measured by multiparameter flow cytometry. Double antibody sandwich enzyme-linked immuno sorbent assay ELISA (DAS-ELISA) was used for detecting the levels of IL-6, IL-10, and IFN-γ. RESULTS: Among the 78 elderly EC patients, CTC level after the surgery was higher than that during the surgery, and CTC level during surgery was higher than that before the surgery (both P < 0.05). Postoperative CTC level in the MIE group was lower than that in the OE group, and the variation of CTC level from pre-operation to intra-operation in the MIE group was also lower than that in the OE group (both P < 0.05). Furthermore, there was significant difference in the incidences of intra-operative and postoperative complications between the MIE group and the OE group (17 cases vs. 31 cases, P < 0.05), and the CTC levels of the patients with complications in either group were significantly higher than the patients without complications (both P < 0.05). IL-6 and IL-10 levels significantly increased, while IFN-γ level decreased in both groups during the surgery and 3 days after the surgery compared to those before the surgery; 2 weeks after the surgery, IL-6 and IL-10 levels in the MIE group recovered to the pre-operative levels (all P < 0.05). However, in the OE group, IL-6 and IL-10 levels 2 weeks after the surgery were still significantly higher than those before the surgery (all P < 0.05); IFN-γ levels in both groups recovered to the pre-operative levels, with higher level in the MIE group than that in the OE group (P < 0.05). CONCLUSION: MIE helped to reduce the survival rate of tumor cells in peripheral blood at the early period of postoperation, and dynamic monitoring CTC level could be used to evaluate the prognosis of EC patients.

Correlations of Promoter Methylation in WIF-1, RASSF1A, and CDH13 Genes with the Risk and Prognosis of Esophageal Cancer.

BACKGROUND This study was designed to explore the correlations of promoter methylation in Wnt inhibitory factor-1 (WIF-1), ras-association domain family member 1A (RASSF1A), and Cadherin 13 (CDH13) genes with the risk and prognosis of esophageal cancer (EC). MATERIAL AND METHODS A total of 71 EC tissues from resection and 35 adjacent normal tissues were collected. Methylation status in the promoter region was detected by methylation- and non-methylation-specific primers. Corresponding mRNA levels were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Correlations between the methylations of these 3 genes and clinicopathologic characteristics were analyzed. Kaplan-Meier method and Cox regression model were used to investigate the relationships between WIF-1, RASSF1A, and CDH13 promoter methylations and the prognosis of EC. RESULTS Compared with adjacent normal tissues, the methylation frequencies of WIF-1, RASSF1A, and CDH13 genes were significantly higher but the mRNA levels of these 3 genes were significantly lower in EC tissues (all P<0.05). WIF-1 and CDH13 promoter methylations were associated with the degree of tumor differentiation and WIF-1 and RASSF1A promoter methylations were associated with age (all P<0.05). The survival rates of patients with WIF-1, RASSF1A, and CDH13 methylations were significantly lower than those of patients without methylation (all P<0.05). WIF-1, RASSF1A, and CDH13 promoter methylations were independent risk factors affecting the prognosis of EC (all P<0.05). CONCLUSIONS WIF-1, RASSF1A, and CDH13 promoter methylations are associated with EC. The methylation levels are negatively related with the prognosis in EC.

The Higher Expression of CDCA2 Associated with Poor Prognosis in Glioma.

Glioma is the most common primary intracranial tumor and is related to poor clinical outcomes. The developments of sensitive markers can be applied to reveal the mechanisms involved in the progression of glioma. This study examined CDCA2 expression in glioma samples and its significance in predicting glioma patient outcome. GEPIA and GEO datasets were used to explore the expression of CDCA2 in glioma. Kaplan-Meier and multivariate assays were applied to delve into the prognostic values of CDCA2 expression in glioma patients using CGGA datasets. Our group also determined the associations between CDCA2 and clinical characteristics. Coexpression analysis was performed. In this research, we observed that CDCA2 expression was distinctly upregulated in glioma specimens compared with nontumor specimens. The prognosis of glioma with high CDCA2 expression was distinctly worse compared with that of glioma with low CDCA2 expression. Additionally, multivariate Cox regression analysis revealed that high CDCA2 expression was an independent poor prognostic indicator for glioma patients. High expression of CDCA2 was positively associated with advanced clinical progression. Coexpression analysis revealed that CDCA2 could be positively related to ASPM, SKA1, DLGAP5, NCAPG, and CDCA8 and was negatively associated with ETNPPL, LDHD, MRVI1, CBX7, and CENPJ. Overall, our findings revealed that CDCA2 might serve as an independent prognosis indicator for glioma.

TTDA inhibited apoptosis by regulating the p53-Bax/Bcl2 axis in glioma.

The trichothiodystrophy group A protein (TTDA) functions in nucleotide excision repair and basal transcription. TTDA plays a role in cancers and serves as a prognostic and predictive factor in high-grade serous ovarian cancer; however, its role in human glioma remains unknown. Here, we found that TTDA was overexpressed in glioma tissues. In vitro experiments revealed that TTDA overexpression inhibited apoptosis of glioma cells and promoted cell growth, whereas knockdown of TTDA had the opposite effect. Increased TTDA expression significantly decreased the Bax/Bcl2 ratio and the level of cleaved-caspase3. TTDA interacted with the p53 gene at the -1959 bp and -1530 bp region and regulated its transcription, leading to inhibition of the p53-Bax/Bcl2 mitochondrial apoptosis pathway in glioma cells. These results indicate that TTDA is an upstream regulator of p53-mediated apoptosis and acts as an oncogene, suggesting its value as a potential molecular target for the diagnosis and treatment of glioma.

In vivo and in vitro effects of hyperplasia suppressor gene on the proliferation and apoptosis of lung adenocarcinoma A549 cells.

Lung adenocarcinoma is the most common subtype of non-small cell lung cancer (NSCLC). Hyperplasia suppressor gene (HSG) has been reported to inhibit cell proliferation, migration, and remodeling in cardiovascular diseases. However, there lacks systematic researches on the effect of HSG on the apoptosis and proliferation of lung adenocarcinoma A549 cells and data of in vivo experiments. The present study aims to investigate the effects of HSG gene silencing on proliferation and apoptosis of lung adenocarcinoma A549 cells. The human lung adenocarcinoma A549 cell was selected to construct adenovirus vector. Reverse transcription-quantitative PCR (RT-qPCR) and Western blot analysis were conducted to detect expressions of HSG and apoptosis related-proteins. Cell Counting Kit (CCK)-8 assay was performed to assess A549 cell proliferation and flow cytometry to analyze cell cycle and apoptosis rate. The BALB/C nude mice were collected to establish xenograft model. Silenced HSG showed decreased mRNA and protein expressions of HSG, and elevated A549 cell survival rates at the time point of 24, 48, and 72 h. The ratio of cells at G0/G1 phase and apoptosis rate decreased and the ratio of cells at S- and G2/M phases increased following the silencing of HSG. There were decreases of B cell lymphoma-2 (Bcl-2)-associated X protein (Bax), Caspase-3, and Caspase-8 expressions but increases in Bcl-2 induced by silenced HSG. As for the xenograft in nude mice, tumor volume increased, and apoptosis index (AI) decreased after HSG silencing. These results indicate that HSG gene silencing may promote the proliferation of A549 cells and inhibit the apoptosis. HSG may be a promising target for the treatment of lung adenocarcinoma.

Association of Mixed Lineage Kinase Domain-Like Protein Expression With Prognosis in Patients With Colon Cancer.

BACKGROUND: The mixed lineage kinase domain-like protein has recently been identified as a key downstream component of tumor necrosis factor-induced necroptosis, which is an important pathway of cancer cell death. The goal of the current study is to explore the expression of mixed lineage kinase domain-like protein in colon cancer tissues and evaluate the prognostic value in patients with colon cancer. METHODS: We collected normal and cancer colon tissues from 135 patients diagnosed with colon cancer after radical operation during July 2007 to April 2009 at The Affiliated Hospital of Qingdao University. Immunohistochemistry analysis was scored using an established scoring system. Kaplan-Meier survival curves were generated for recurrence-free survival and overall survival for all patients and 2 subsets of patients. The relationship between mixed lineage kinase domain-like protein expression and prognosis parameter (recurrence-free survival, overall survival) was analyzed by univariate and multivariate Cox regression analyses. RESULTS: The median age of all patients was 67 years and 56.3% were male. Low expression of mixed lineage kinase domain-like protein was associated with decreased overall survival (78.6 vs 81.2 months; P = .011) in all patients. In the subset of 79 patients who received adjuvant chemotherapy, low expression of mixed lineage kinase domain-like protein was associated with decreased recurrence-free survival (60.4 vs 72.8 months; P = .032) and decreased overall survival (66.3 vs 72.9 months; P = .005). Low expression of mixed lineage kinase domain-like protein was associated with decreased overall survival (74.9 vs 79.8 months; P = .006) and recurrence-free survival (69.6 vs 78.8 months; P = .005) among patients with Tumor Node Metastasis (TNM) stage II colon cancer. CONCLUSIONS: Low expression of mixed lineage kinase domain-like protein was associated with decreased overall survival in all patient-group with resected colon cancer. It is associated with decreased recurrence-free survival and overall survival in the subset of patients who receive adjuvant chemotherapy and patients who were TNM stage II. Mixed lineage kinase domain-like protein may provide important prognostic information in patients with colon cancer.

DNA hypermethylation of the vimentin gene inversely correlates with vimentin expression in intestinal- and diffuse-type gastric cancer.

The vimentin gene is a hallmark of epithelial-to-mesenchymal transition and has been observed to be overexpressed in various types of tumor cell line and tissue. Previous studies have reported correlations between vimentin DNA methylation levels and subsequent vimentin expression levels in solid tumors, including breast and colorectal cancer; however, to the best of our knowledge, such a correlation has not been reported for gastric cancer (GC) using Lauren classification. Therefore, the present study aimed to quantify DNA methylation levels of the vimentin gene using quantitative (q) methylation-specific polymerase chain reaction (PCR) in intestinal-type GC cell lines (MKN-28, AGS and MKN-1), diffuse-type GC cell lines (SGC-7901, SNU-5 and KATO III), the GES-1 immortalized human non-neoplastic gastric epithelial cell line, as well as in tumor and paratumor normal tissue samples. Furthermore, the present study analyzed the messenger RNA expression of the vimentin gene in these cell lines and tissues by reverse transcription-qPCR. A comparison of the clinicopathological features was conducted between patients, grouped according to the Lauren classification. The present study identified that the vimentin promoter region was hypermethylated in all GC cell lines and tumor tissue samples when compared with immortalized normal gastric epithelial cells and paratumor normal tissues. In addition, vimentin promoter methylation levels were observed to be higher in intestinal-type cell lines when compared with those of diffuse-type lines and tissues. Correspondingly, vimentin expression levels were lower in intestinal-type gastric cell lines compared with those of diffuse-type cell lines and tissues, and were lowest in the non-neoplastic gastric cell line and paratumor normal tissues. Patients with diffuse-type GC were on average younger (P=0.023), and exhibited higher tumor (P=0.020), node (P=0.032) and TNM classification of malignant tumor stage (P=0.039) than those with intestinal-type GC. Following treatment of AGS cells (which demonstrated the highest methylation level of the vimentin gene) with 5-aza-2'-deoxycytidine, vimentin expression was restored significantly. Thus, the present study revealed that vimentin promoter methylation levels are inversely correlated with vimentin expression levels in GC (according to Lauren classification). High levels of methylation in the vimentin gene promoter region may be involved in carcinogenesis and the development of GC, and may provide a novel molecular classification for GC.

[A preliminary study of stenting followed by laparoscopic surgery for obstructing left-sided colon cancer].

OBJECTIVE: To study the efficacy of stenting followed by laparoscopic surgery in the treatment of obstructing left-sided colon cancer. METHODS: Forty-nine patients with obstructing left-sided colon cancer were prospectively randomized into two groups. Twenty patients received emergent open surgery, while 15 underwent laparoscopic surgery 3 days after placement of the self-expanding metal stent (SEMS) and 14 of them received laparoscopic surgery 10 days after placement of SEMS. Outcomes evaluated included 1-stage operation rate, conversion rate, operative time, length of hospital stay, blood loss, postoperative pain score and use of analgesics, rates of permanent stoma, and postoperative complications. RESULTS: Compared with emergent open surgery, patients undergoing laparoscopic surgery had significantly less blood loss(P=0.000), lower permanent stoma rate (P=0.024), less pain(P=0.000), and lower incidence of postoperative complications. Laparoscopic surgery was associated with a significantly higher rate of 1-stage operation(P=0.004). Compared with patients undergoing laparoscopic surgery 3 days after SEMS placement, patients who underwent laparoscopic surgery 10 days after SEMS placement had a significantly higher 1-stage operation rate(P=0.001) and a lower conversion rate(P=0.046). CONCLUSIONS: Self-expanding metal stenting is a safe and effective bridge to laparoscopic surgery in patients with obstructing left-sided colon cancer. Laparoscopic surgery 10 days after SEMS placement may be more appropriate.

[Study on No.7 lymph nodes micrometastasis in patients with node- negative gastric carcinoma by routine examination].

OBJECTIVE: To investigate the micrometastatic characteristics in the gastric cancer patients with negative lymph node invasion by routine pathological examination and evaluate the relationship between micrometastasis and clinicopathological features. METHODS: One hundred and thirty-eight lymph nodes from No. 7 group in 46 patients with node-negative gastric cancer by routine examination were examined by consecutive sections. Telomerase activity was determined by RT-PCR and ELISA. Clinicopathological data were analyzed by statistical method. RESULTS: Micrometastasis was found in 32 lymph nodes (23.2%) from 13 cases (28.3%) by consecutive sections. Telomerase activity was positive in 49 lymph nodes (35.5%) from 20 cases (43.5%). The sensitivity, specificity, positive and negative predictive value and diagnostic accuracy of TRAP-ELISA was 100%, 84%, 65%, 100% and 88% respectively. The micrometastasis in lymph node was correlated with age, sex, primary tumor location, histological classification and metastatic lymph node type (P> 0.05), but correlated with the gross type of the primary tumor, size and serosa invasion (P< 0.05). CONCLUSIONS: It is necessary to discern the micrometastasis in gastric cancer with negative lymph node by routine examination in order to objectively evaluate the clinicopathological classification and prognosis. TRAP-ELISA can be a complementary method to traditional histological examination.