FTO-mediated ZNF687 accelerates tumor growth, metastasis, and angiogenesis in colorectal cancer through the Wnt/ß-catenin pathway.

Colorectal cancer (CRC) is a common and lethal cancer. ZNF687 has been disclosed to take part in diversified cancers' progression by serving as a facilitator. However, the detailed regulatory functions of ZNF687 in the CRC have not been investigated. This work is planned to probe the impacts of ZNF687 on CRC progression. The IHC, RT-qPCR, and western blot assays were used to examine mRNA and protein gene expressions. The cell proliferation measurement was accompanied by a CCK-8 assay. The Transwell assay was performed to evaluate cell invasion and migration. The angiogenesis ability was evaluated by a tube formation experiment. The m6A level was evaluated through MeRIP and m6A dot blot assays. The binding ability between ZNF687 and FTO (fat mass and obesity associated protein) was tested through an RIP assay. The ß-catenin nuclear translocation was assessed through an immunofluorescence assay. The tumor growth was evaluated through an in vivo assay. ZNF687 exhibited higher expression in CRC cells and resulted in a poor prognosis. Additionally, ZNF687 inhibition suppressed CRC cell proliferation, invasion, migration, and angiogenesis. Furthermore, the suppression of ZNF687 retarded the Wnt pathway. Through rescue assays, the reduced cell migration, proliferation, invasion, and angiogenesis mediated by ZNF687 knockdown could be reversed after BML-284 (the activator of the Wnt pathway) treatment. Finally, it was explained that ZNF687 knockdown inhibited in vivo tumor growth. This study manifested that FTO-mediated ZNF687 aggravated tumor growth, metastasis, and angiogenesis of CRC through Wnt/ß-catenin pathway. This finding may provide a hopeful molecular target for CRC treatment.

Microarray profile analysis identifies ETS1 as potential biomarker regulated by miR-23b and modulates TCF4 in gastric cancer.

BACKGROUND: Gastric cancer (GC), a common malignancy of the human digestive system, represents the second leading cause of cancer-related deaths worldwide. Early detection of GC has a significant impact on clinical outcomes. The aim of this study was to identify potential GC biomarkers. METHODS: In this study, we conducted a multi-step analysis of expression profiles in GC clinical samples downloaded from TCGA database to identify differentially expressed miRNAs (DEMs) and differentially expressed mRNAs (DEGs). Potential prognostic biomarkers from the available DEMs were then established using the Cox regression method. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to investigate the biological role of the predicted target genes of the miRNA biomarkers. Then, the prognostic DEM-mediated regulatory network was constructed based on transcription factor (TF)-miRNA-target interaction. Subsequently, the consensus genes were further determined based on the overlap between DEGs and these target genes of DEMs. Besides, expression profile, co-expression analysis, immunity, and prognostic values of these prognostic genes were also investigated to further explore the roles in the mechanism of GC tumorigenesis. RESULTS: We got five miRNAs, including miR-23b, miR-100, miR-143, miR-145, and miR-409, which are associated with the overall survival of GC patients. Subsequently, enrichment analysis of the target genes of the miRNA biomarkers shown that the GO biological process terms were mainly enriched in mRNA catabolic process, nuclear chromatin, and RNA binding. In addition, the KEGG pathways were significantly enriched in fatty acid metabolism, extracellular matrix (ECM) receptor interaction, and proteoglycans in cancer pathways. The transcriptional regulatory network consisting of 68 TFs, 4 DEMs, and 58 targets was constructed based on the interaction of TFs, miRNAs, and targets. The downstream gene ETS1 of miR-23b and TCF4 regulated by ETS1 were obtained by the regulatory network construction and co-expression analysis. High expression of ETS1 and TCF4 indicated poor prognosis in GC patients, particularly in the advanced stages. The expression of ETS1 and TCF4 was correlated with CD4+ T cells, CD8+ T cells, and B cells. CONCLUSIONS: miR-23b, ETS1, and TCF4 were identified as the prognostic biomarkers. ETS1 and TCF4 had potential immune function in GC, which provided a theoretical basis for molecular-targeted combined immunotherapy in the future.

Elevated Neutrophil-to-Lymphocyte Ratio Is Associated With Poor Outcomes for Melanoma Patients Treated With PD-1 Inhibitor or Chemotherapy in a Chinese Population.

Background: Previous studies have suggested that an elevated pre-treatment neutrophil-to-lymphocyte ratio (NLR) is associated with worse outcomes in patients with a variety of cancers. The purpose of this retrospective analysis is to investigate the prognostic value of the NLR in a Chinese melanoma population. Methods: Melanoma patients were divided into two groups based on pre-treatment NLR values (≥3 vs. <3). Cox proportional hazard regression analysis and the Kaplan-Meier method were employed to study the prognostic role of the NLR for overall survival (OS) and progression-free survival (PFS). Results: A total of 159 melanoma patients were included in this study, including 40 patients treated with PD-1 inhibitor and 119 patients treated with chemotherapy. In the PD-1 inhibitor group, the median OS was 18.0 months in the low NLR subgroup and 5.6 months in the high NLR subgroup; the median PFS was 7.0 and 2.2 months, respectively. In chemotherapy group, the median OS was 23.0 months in the low NLR group and 8.0 months in the high NLR group, and the median PFS was 9.0 and 4.0 months, respectively. Multivariate analysis showed that the NLR was significantly associated with OS and PFS in melanoma patients treated with either PD-1 inhibitor immunotherapy or chemotherapy. Conclusion: In the Chinese population, an elevated NLR was closely related to worse survival in patients with melanoma treated with either PD-1 inhibitor monotherapy or chemotherapy.

Elevated platelet-to-lymphocyte corresponds with poor outcome in patients with advanced cancer receiving anti-PD-1 therapy.

OBJECTIVES: The purpose of this retrospective analysis was to investigate the prognostic value of PLR for PD-1 inhibitors. METHODS: Patients were divided into different subgroups according to PLR. Univariate survival analysis and a multivariate Cox proportional hazards regression model were used to assess the association between PLR and overall survival (OS) or progression-free survival (PFS). RESULTS: The optimal cut-off value of baseline PLR was 164. Among the total 85 patients, 34 patients presented with PLR ≥ 164, and 51 presented with PLR < 164, respectively. The median OS for the high PLR group was 7.0 months (95% CI: 4.1-9.9 months), and it was not reached for the low PLR group (P < 0.001). The median PFS was 3.0 months (95% CI: 1.9-4.1 months) vs. 9.8 months (95% CI: 6.1-13.5 months) for the high and low PLR groups, respectively (P < 0.001). In multivariate analysis, a PLR > 164 and body mass index (BMI) > 24.0 were independently associated with OS (hazard ratio [HR]: 3.549, 95% confidence interval [CI]: 1.901-6.625, P < 0.001 and HR: 0.496, 95% CI: 0.260-0.945, P = 0.033), meanwhile PLR was also significantly associated with inferior PFS (HR: 2.567, 95% CI: 1.551-4.249, P < 0.001). Disease control rate for high and low PLR group was 38.2% and 74.5%, respectively, and it was also correlated with elevated PLR (P = 0.001). CONCLUSION: This retrospective analysis indicates that PLR could be used as a biomarker to stratify patients who will have a better response to anti-PD-1 agents.

Advanced colorectal adenoma related gene expression signature may predict prognostic for colorectal cancer patients with adenoma-carcinoma sequence.

BACKGROUND: There are still no absolute parameters predicting progression of adenoma into cancer. The present study aimed to characterize functional differences on the multistep carcinogenetic process from the adenoma-carcinoma sequence. METHODS: All samples were collected and mRNA expression profiling was performed by using Agilent Microarray high-throughput gene-chip technology. Then, the characteristics of mRNA expression profiles of adenoma-carcinoma sequence were described with bioinformatics software, and we analyzed the relationship between gene expression profiles of adenoma-adenocarcinoma sequence and clinical prognosis of colorectal cancer. RESULTS: The mRNA expressions of adenoma-carcinoma sequence were significantly different between high-grade intraepithelial neoplasia group and adenocarcinoma group. The biological process of gene ontology function enrichment analysis on differentially expressed genes between high-grade intraepithelial neoplasia group and adenocarcinoma group showed that genes enriched in the extracellular structure organization, skeletal system development, biological adhesion and itself regulated growth regulation, with the P value after FDR correction of less than 0.05. In addition, IPR-related protein mainly focused on the insulin-like growth factor binding proteins. CONCLUSION: The variable trends of gene expression profiles for adenoma-carcinoma sequence were mainly concentrated in high-grade intraepithelial neoplasia and adenocarcinoma. The differentially expressed genes are significantly correlated between high-grade intraepithelial neoplasia group and adenocarcinoma group. Bioinformatics analysis is an effective way to study the gene expression profiles in the adenoma-carcinoma sequence, and may provide an effective tool to involve colorectal cancer research strategy into colorectal adenoma or advanced adenoma.

Elevated microRNA-23a Expression Enhances the Chemoresistance of Colorectal Cancer Cells with Microsatellite Instability to 5-Fluorouracil by Directly Targeting ABCF1.

Colorectal cancer (CRC) is one of the most common tumor types worldwide. A frequent subtype of CRC is defined by a deficiency in the mismatch repair (MMR) pathway, constantly found in combination with microsatellite instability (MSI), which not only contributes to the pathogenesis of a large proportion of CRC, but also controls the response to multiple drugs used to treat CRCs. The most commonly used chemotherapeutic agent for CRC is 5-fluorouracil (5-FU). However, CRC with MSI frequently acquires 5-FU resistance, and the exact mechanism underlying how CRC cells acquire chemoresistance to 5-FU remains incompletely understood. Recently, emerging evidence has demonstrated that microRNAs (miRNAs) are key players in multidrug resistance. In this study, we aimed to characterize the expression profiles and functions of miRNAs in 5-FU-resistant CRC with MSI. We found that miR-23a was significantly elevated in MSI CRC cells and tissues compared to microsatellite stability (MSS) CRC cells and tissues. Ectopic expression of miR-23a increased the viability and survival of MSS CRC cells. Inversely, downregulation of miR-23a reduced viability in and promoted cell apoptosis in MSI CRC cells treated with 5-FU. Moreover, we demonstrated that ABCF1 is a direct target of miR-23a. Additionally, the expression of miR-23a was inversely correlated with the expression of ABCF1 in CRC tissues. Interestingly, repressing ABCF1 expression by either miR-23a overexpression or siABCF1 led to recovery of 5-FU sensitivity in MSI CRC cells. These data demonstrated that miR-23a enhances 5-FU resistance in MSI CRC cells through targeting ABCF1 and thus provided important implications for therapeutic approaches aiming to overcome MSI CRC resistance to 5-FU.

Surgical management of leiomyosarcoma of the inferior vena cava.

The optimal surgical management of patients with leiomyosarcoma of inferior vena cava remains a controversy. From 1975 and 2009, five patients with leiomyosarcoma of inferior vena cava were treated at the Chinese PLA General Hospital and Beijing Shijitan Hospital. The age ranged 39-61 years and the duration of symptoms ranged from 18 to 36 months. Abdominal and back pain are the most common complaints. A combination of various imaging modalities is essential for treatment planning. R0, R1, R2, and biopsy only were accomplished in 2, 1, 1, and 1 case, respectively. Combined resections included inferior vena cava, right kidney, adrenal gland, psoas, colon, duodenal, gallbladder, liver, and/or aorta, without inferior vena cava reconstruction. No inferior vena cava-related postoperative complication was seen in our series.

Two Chinese pedigrees for adenomatous polyposis coli: new mutations at codon 1309 and predisposition to phenotypic variations.

Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease caused by a mutation in the adenomatous polyposis coli (APC) gene. Some studies have attempted to correlate mutations at codon 1309 with classic FAP (≥100 colorectal polyps). We report two Chinese FAP pedigrees with new frameshift mutations at codon 1309, in which affected individuals manifest phenotypic variations. Comprehensive physical examinations were performed for all living individuals and the medical data of deceased patients were collected. Screening of the APC and human mutY homolog (MUTYH) genes for germline mutations was conducted by direct polymerase chain reaction (PCR) sequencing. In two pedigrees, a heterozygous deletion in exon 16 of the APC gene was present in all FAP patients but absent in the unaffected individuals. There were no changes to the MUTYH gene. The first pedigree, with a new frameshift mutation at c.3926_3930 del AAAAG (p. Glu1309Aspfs X4), exhibited obvious differences in the polyp number such that the proband manifested only three colorectal polyps, whereas another patients showed the symptoms of classic FAP. The second pedigree, also traced a new mutation at c.3922_3925 del AAAG (p. Glu1309Argfs X11). Although all of the patients presented with classic polyposis, one of them exhibited a delayed onset of colorectal cancer in his 50s. Two novel mutations at codon 1309 in two Chinese families suffering from FAP could enrich the germline mutation spectrum of the APC gene. Families of individuals might manifest different phenotypes, even with an identical codon 1309 mutation, unlike in previous studies.

Salvage irrigation-suction in gracilis muscle repair of complex rectovaginal and rectourethral fistulas.

AIM: To evaluate the efficacy of gracilis muscle transposition and postoperative salvage irrigation-suction in the treatment of complex rectovaginal fistulas (RVFs) and rectourethral fistulas (RUFs). METHODS: Between May 2009 and March 2012, 11 female patients with complex RVFs and 8 male patients with RUFs were prospectively enrolled. Gracilis muscle transposition was undertaken in all patients and postoperative wound irrigation-suction was performed in patients with early leakage. Efficacy was assessed in terms of the success rate and surgical complications. SF-36 quality of life (QOL) scores and Wexner fecal incontinence scores were compared before and after surgery. RESULTS: The fistulas healed in 14 patients after gracilis muscle transposition; the initial healing rate was 73.7%. Postoperative leakage occurred and continuous irrigation-suction of wounds was undertaken in 5 patients: 4 healed and 1 failed, and postoperative fecal diversions were performed for the patient whose treatment failed. At a median follow-up of 17 mo, the overall healing rate was 94.7%. Postoperative complications occurred in 4 cases. Significant improvement was observed in the quality outcomes framework scores (P < 0.001) and Wexner fecal incontinence scores (P = 0.002) after the successful healing of complex RVFs or RUFs. There was no significant difference in SF-36 QOL scores between the initial healing group and irrigation-suction-assisted healing group. CONCLUSION: Gracilis muscle transposition and postoperative salvage wound irrigation-suction gained a high success rate in the treatment of complex RVFs and RUFs. QOL and fecal incontinence were significantly improved after the successful healing of RVFs and RUFs.

[Prospective study of gracilis muscle repair of complex rectovaginal fistula and rectourethral fistula].

OBJECTIVE: To assess the efficacy and experience of gracilis muscle transposition for complex rectovaginal fistula (RVF) and rectourethral fistula (RUF). METHODS: Nineteen patients underwent gracilis muscle transposition for complex RVF and RUF from May 2009 to November 2011 in the Beijing Shijitan Hospital and the clinical data were prospectively collected. The success rate and complications were recorded. SF-36 quality of life score, Wexner fecal incontinence score, and female sexual function score before surgery and 6 months after surgery were recorded. RESULTS: In 19 patients, there were 8 males (RUF) and 11 females (RUF). The times of failed attempt repair preoperatively ranged from 0-3 (mean, 1.0). The diameter of the fistula ranged from 0.5-3.0 cm (mean, 1.6), and all fistulas located above the sphincter. The operative time ranged from 145-400 minutes (median, 240). The postoperative hospital stay ranged from 10-39 days (median 21). Early postoperative complications included thigh pain and numbness in 2 cases, leg numbness in 2 cases. No long-term complications were noticed. The follow-up period ranged from 6-35 months (median, 18). The gracilis muscle transposition had a healing rate of 94.7% (18/19). As compared with the preoperative level, Wexner score decreased from 10.0±8.8 to 2.9±5.8, and the continence function improved significantly (P=0.002). Sexual function score of 11 female patients increased from 1.0±1.8 to 4.0±4.0, and the sexual function had a significant improvement after surgery (P=0.022). SF-36 quality of life scores improved significantly (P<0.001). CONCLUSIONS: Gracilis muscle transposition for complex rectovaginal fistula and rectourethral fistula has high success rate with mild and rare complications.

[Bloodless medical and surgical procedures for retroperitoneal neoplasm resection].

OBJECTIVE: To investigate the Bloodless Medical and Surgical Procedures for retroperitoneal neoplasm resection. METHODS: Retrospectively analyse the Bloodless Medical and Surgical Procedures during 36 retroperitoneal neoplasm resections from Beijing Shijitan Hospital Affiliated to Capital Medical University from September 2009 to December 2010, to discuss the preoperative preparation, anesthetic induction and maintainance, intraoperative monitoring and use of vasoactive drugs related experience. RESULTS: All the patients were safe during the perioperative period, without any operative and anesthetic complication. By applying the Bloodless Medical and Surgical Procedures, the intraoperative haemodynamics maintained steady, the mean hematocrit decreased from 0.368 ± 0.095 before autologous blood collection to 0.252 ± 0.032 before the ends of operation. Majority of the patients (91.7%) stop using vasoactive drugs and extubated within 1 h after operation, and return wards. CONCLUSION: Erythrocyte-raising medicine therapy and modified preservation autologous blood transfusion are important process of Bloodless Medical and Surgical Procedures.

[Expression and clinical significance of LRP16 gene in human breast cancer].

BACKGROUND & OBJECTIVE: Estrogen directly up-regulates LRP16 gene expression via activating its receptor (ER), and the overexpression of LRP16 promotes the proliferation of human breast cancer cells. This study was to detect the mRNA level of LRP16 gene in breast cancer, and investigate its correlation to the clinicopathologic features. METHODS: The mRNA level of LRP16 in carcinoma and matched peritumor tissues from 22 breast cancer patients was detected by Northern blot, and that in the tissues from 30 patients was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The expression of Ki-67, ER, and progesterone receptor (PR) in the carcinoma tissues was detected by immunohistochemistry. RESULTS: According to the results of Northern blot, compared with that in peritumor tissues, LRP16 was overexpressed by 2 folds in 9 (40.9%) out of 22 breast cancer samples. Of the 9 samples with LRP16 overexpression, 7 were ER-positive, and 8 were PR-positive; of the 13 samples without LRP16 overexpression, 6 were ER-positive, and 5 were PR-negative. The positive rates of ER and PR were significantly higher in the samples with LRP16 overexpression than in the samples without LRP16 overexpression (P<0.05). Only 1 of the 9 samples with LRP16 overexpression was negative for both ER and PR, but 7 of the 13 without LRP16 overexpression were negative for both of them. The proportion of the tumors with diameters of 3.0-4.5 cm was significantly higher in the patients with LRP16 overexpression than in those without LRP16 overexpression (8/9 vs. 5/13, P=0.031). Axillary lymph node metastasis was detected in 12 out of 22 patients, including 8 of the 9 patients with LRP16 overexpression and 4 of the 13 without LRP16 overexpression (P=0.011). In addition, LRP16 overexpression was detected in 6 of the 8 patients with Ki-67 overexpression, and 2 of the 14 patients without Ki-67 overexpression (P=0.026). According to the results of RT-PCR, LRP16 was overexpressed in 9 (30%) out of 30 breast cancer samples. All of the 9 samples with LRP16 overexpression were positive for both ER and PR, with Ki-67 overexpression, tumor diameters of more than 3.5 cm and axillary lymph node metastasis. The differences between the patients with or without LRP16 overexpression were significant (P<0.05). CONCLUSION: LRP16 overexpression is closely correlated to the positive rates of ER and PR, Ki-67 level, tumor diameter, and axillary lymph node metastasis of breast cancer, and might be involved in the proliferation and metastasis of human breast cancer.