CHD1 deletion stabilizes HIF1α to promote angiogenesis and glycolysis in prostate cancer / 亚洲男科学杂志(英文版)

Chromodomain-helicase-DNA-binding protein 1 (CHD1) deletion is among the most common mutations in prostate cancer (PCa), but its role remains unclear. In this study, RNA sequencing was conducted in PCa cells after clustered regularly interspaced palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9)-based CHD1 knockout. Gene set enrichment analysis (GSEA) indicated upregulation of hypoxia-related pathways. A subsequent study confirmed that CHD1 deletion significantly upregulated hypoxia-inducible factor 1α (HIF1α) expression. Mechanistic investigation revealed that CHD1 deletion upregulated HIF1α by transcriptionally downregulating prolyl hydroxylase domain protein 2 (PHD2), a prolyl hydroxylase catalyzing the hydroxylation of HIF1α and thus promoting its degradation by the E3 ligase von Hippel-Lindau tumor suppressor (VHL). Functional analysis showed that CHD1 deletion promoted angiogenesis and glycolysis, possibly through HIF1α target genes. Taken together, these findings indicate that CHD1 deletion enhances HIF1α expression through PHD2 downregulation and therefore promotes angiogenesis and metabolic reprogramming in PCa.

Annexin A7 inhibits proliferation of hepatoma cells via binding with IGFBP2 / 第二军医大学学报

Objective To investigate the role of Annexin A7CANXA7) in the development of HCC by analyzing ANXA7 expression in hepatocarcinogenesis and identifying its potential interaction molecule. Methods ANXA7 mRNA expression was analyzed by real-time PCR in 48 HCC tissues and tumor adjacent tissues, and different hepatic cancer tissues and cell lines. To analyze the effect of ANXA7 on hepatoma proliferation, ANXA7 was overexpressed or inhibited by specific siRNA in hepatic cancer cells. Co-immunopricipitation (co-IP) method was used to detect the specific binding protein of ANXA7 in HCC cells. The key sites of protein interaction were analyzed by point mutation. Western blotting analysis was used to study the effect of ANXA7 on IGFBP2 activated ERK1/2 phosphorylation. Results The expression of ANXA7 was down-regulated in both hepatoma tissue samples and hepatoma cell lines. Insulin-like growth factors binding protein 2 (IGFBP2) could specifically bind with ANXA7 through the key ROD site. Up-regulated expression of ANXA7 could inhibit the proliferation of tumor cells (P

Expression and purification of AnxB1-MLT chimera and its inhibitory effect on SMMC7721 and HepG2 cell proliferation / 第二军医大学学报

Objective To obtain a chimera composed of annexin Bl (AnxBl) and melittin (MLT) and to investigate its inhibitory effect on phosphatidylserine liposome activity and SMMC7721 and HepG2 cell proliferation. Methods A fusion gene AnxBl-MLT was constructed by overlap extension gene splicing and then was inserted into plasmid pGEX-5T. The recombinant plasmid was transformed into E. coli strain K802 and induced by IPTG at low temperature. The expression condition was optimized and GST affinity chromatography column was used for purification. Calcium-dependent phospholipid binding assay was used to determine whether the chimera kept the activity of AnxBl. CCK8 analysis was employed to investigate the effect of AnxBl-MLT on SMMC7721 and HepG2 cell proliferation. Results After being inserted into the expression plasmid, AnxBl-MLT protein expressed in K802 cells, with a high level recombinant protein induced by 0. 2 mmol/ L IPTG at 22-24»C for 4 h. The protein AnxBl-MLT was purified by using GST affinity chromatography column (a band at 63 000) and the purification of the final purified protein was >95%. AnxBl-MLT was able to bind to phosphatidylserine liposome in a calcium-dependent manner. CCK8 analysis indicated that AnxBl-MLT inhibited SMMC7721 and HepG2 cell proliferation at a dose-dependent manner. Conclusion We have successfully constructed a chimera AnxBl-MLT, which retains the calcium-dependent phospholipid binding activity of AnxB1, and can inhibit the proliferation of hepatic cancer cell lines SMMC7721 and HepG2.

Diagnostic accuracy research of needle puncture biopsy during operation for pulmonary single nodules / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the diagnostic accuracy of needle puncture biopsy and pathological examination of frozen during operation for pulmonary nodules, and whether this diagnostic method can replace tumor resection examination.</p><p><b>METHODS</b>Totally 50 patients (28 males and 22 females, average age was 59 years) who had the single nodule after imaging examination without any pathological diagnostic from January to October 2010 were selected in this research work. During open operation or video assisted thoracic surgery, needle (14 G model) was used to puncture biopsy for pathological examination of frozen. All the adverse events during puncture biopsy would be recorded. The resection specimens would be accepted paraffin pathological examination. The relationship between puncture frozen pathological and paraffin pathological examination was analyzed.</p><p><b>RESULTS</b>All tumor sizes were ranged from 1.0 cm × 0.6 cm to 5.6 cm × 9.0 cm. The paraffin pathological examination after operation as the golden standard, there were 7 cases of benign tumor and 43 cases of malignant tumor. The diagnostic sensitivity of puncture biopsy was 90.7%, the specificity was 100%, the positive predictive value was 100% and the negative predictive value was 63.6%. There were 11 cases of benign tumor diagnosed by needle puncture biopsy, among which 4 cases were proved as malignant tumor by paraffin pathology, and the false negative rate was 9.3%. The main risk of puncture biopsy was bleeding after puncture immediately, and the rate was 4.0% (2/50).</p><p><b>CONCLUSIONS</b>The puncture biopsy during operation had a high specificity for malignant lung tumor, and there was a certain false negative rate for benign tumor. Puncture biopsy and pathological examination of frozen tissue can replace tumor section biopsy in a way.</p>