Colorectal cancer (CRC) is the third most common cancer and causes high mortality worldwide. Although CRC has been studied widely, the molecular mechanism is not completely known. Eukaryotic translation elongation factor 1 delta (EEF1D) participates in the progression of various tumors, however, the effect of EEF1D on CRC remains unclear. Here, we aimed to identify the potential mechanism of EEF1D in CRC. The expression levels of EEF1D were assessed in CRC samples. Functional analysis of EEF1D in CRC was detected in vitro and in vivo. The regulatory mechanism of EEF1D was identified with RNA immunoprecipitation, RNA pull-down assay, and proteomics analysis. Our findings confirmed that EEF1D was upregulated in human CRC tissues. Functionally, EEF1D overexpression accelerated cell proliferation and metastasis, whereas EEF1D knockdown inhibited cell proliferation and metastasis both in vitro and in vivo CRC models. Furthermore, we showed that EEF1D was upregulated by SRSF9 via binding to 3'UTR of EEF1D mRNA. EEF1D knockdown reversed the malignant phenotype induced by SRSF9 overexpression. These findings demonstrated that EEF1D promotes CRC progression, and EEF1D may be a molecular target against CRC.

Ferroptosis, a newly iron-dependent form of cell death, is often accompanied by the damage of membrane lipid peroxide. Recently, the ferroptosis inducer erastin has been reported to exhibit potential anti-cancer activities. The aim of this study was to investigate the effects of SRSF9 on the sensitivity of colorectal cancer (CRC) to erastin and explore the underlying molecular mechanism. Short hairpin RNAs (shRNAs) or SRSF9 overexpression vector (SRSF9-OE) was transfected into erastin-induced human CRC cells to inhibit or overexpress SRSF9. Results showed that SRSF9 inhibition promoted the cell death induced by erastin, conversely, SRSF9 overexpression augmented the resistance to erastin-induced death in human CRC cells. SRSF9 decreased lipid peroxide damage which was a key event during erastin-induced ferroptosis in human CRC cells. Furthermore, we found that SRSF9 inhibition increased erastin-induced ferroptosis by downregulating GPX4 level. In an In vivo study, SRSF9 shRNA or SRSF9-OE stably transfected human CRC cells were subcutaneously injected into the right flank of nude mice. SRSF9 overexpression partly abolished the tumor growth inhibition and ferroptosis induced by erastin. Our data indicated SRSF9's regulation of GPX4 as an essential mechanism driving CRC tumorigenesis and resistance of erastin-induced ferroptosis. This molecular mechanism may provide a novel method for improving the sensitivity of CRC to erastin.

Ferroptosis, a newly discovered form of programmed cell death characterized by lipid peroxidation, crafts a new perspective on cancer treatment. Serine and arginine rich splicing factor 9 (SFRS9) is frequently described as a proto-oncogene in cervical and bladder cancer. However, the role of SFRS9 in colorectal cancer (CRC) and whether SFRS9 exerts its function associated with ferroptosis is largely unknown. Herein, we found that the expression of SFRS9 mRNA and protein in the CRC tissues was obviously higher than that in the paracancerous tissues. Function assays revealed that SFRS9 overexpression (SFRS9-OE) significantly promoted cell viability, cell cycle progression and colony formation of CRC cells. While SFRS9 knockdown by shRNAs transfection inhibited these progressions. Furthermore, cell death and lipid peroxidation induced by ferroptosis inducers erastin and sorafenib were suppressed by SFRS9-OE. Bioinformatics analysis indicated that SFRS9 can bind to peroxidase 4 (GPX4) mRNA which is a central regulator of ferroptosis. Western blot showed that GPX4 protein expression was clearly elevated upon SFRS9-OE, while it was decreased in SFRS9-inhibited CRC cells. RNA immunoprecipitation experiment was carried out in HCT116 cells to confirm the binding of SFRS9 and GPX4 mRNA specifically. SiGPX4 transfection reversed the inhibitory effects of SFRS9-OE on the erastin and sorafenib-induced ferroptosis. Consistent with our in vitro observations, SFRS9 promoted the growth of tumors while SFRS9 knockdown significantly inhibited tumor growth in nude mice. In conclusion, SFRS9 represents an obstructive factor to ferroptosis by upregulating GPX4 protein expression, and knocking down SFRS9 might be an effective treatment for CRC.

Hepatocellular carcinoma (HCC) is the most common visceral neoplasms with its heterogeneity and high rate of recurrence. HCC is characterized to be delayed diagnosis and the development of resistant disease. However, the molecular mechanism for HCC pathogenesis and progression remains largely unknown. Here, we demonstrated that ubiquitin-specific protease14 (USP14) is highly expressed in HCC samples, and the higher expression of USP14 is positively correlated with poor prognosis. Interestingly, USP14 is involved in the maintenance of HIF1-α stability to activate HIF1-α-induced transactivation via its deubiquitinase activity. USP14 depletion or its specific inhibitor IU1 treatment decreased cell proliferation, invasion, migration, and Vascular Mimicry (VM) formation even under hypoxia conditions in HCC cell lines. Moreover, we provided the evidence to show that knockdown of USP14 or USP14 inhibitor (IU1) treatment inhibited tumor growth in tumor-bearing nude mice. Our findings suggest that USP14 maintains HIF1-α stability through its deubiquitination activity, providing a potential biomarker for the early diagnosis and therapy of HCC.

BACKGROUND:Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide, with unmet need for the pharmacological therapy. The functions of ATXN7L3 in HCC progression are not known.METHODS:RNA sequence, quantitative real-time PCR, and western blot were performed to detect gene expression. Chromatin immunoprecipitation was performed to detect possible mechanisms. Immunohistochemical stain was performed to examine the protein expression. Colony formation, cell growth curve and xenograft tumor experiments were performed to examine cell growth in vitro and in vivo.FINDINGS:ATXN7L3 functions as a coactivator for ERα-mediated transactivation in HCC cells, thereby contributing to enhanced SMAD7 transcription. ATXN7L3 is recruited to the promoter regions of SMAD7 gene, thereby regulating histone H2B ubiquitination level, to enhance the transcription of SMAD7. A series of genes regulated by ATXN7L3 were identified. Moreover, ATXN7L3 participates in suppression of tumor growth. In addition, ATXN7L3 is lower expressed in HCC samples, and the lower expression of ATXN7L3 positively correlates with poor clinical outcome in patients with HCC.INTERPRETATION:This study demonstrated that ATXN7L3 is a novel regulator of SMAD7 transcription, subsequently participating in inhibition of tumor growth in HCC, which provides an insight to support a previously unknown role of ATXN7L3 in HCC progression. FUND: This work was funded by 973 Program Grant from the Ministry of Science and Technology of China (2013CB945201), National Natural Science Foundation of China (31871286, 81872015, 31701102, 81702800, 81902889), Foundation for Special Professor of Liaoning Province, Natural Science Foundation of Liaoning Province (No.20180530072); China Postdoctoral Science Foundation (2019M651164).

Androgen receptor (AR) signaling is considered to be crucial for the pathogenesis of hepatocellular carcinoma (HCC) with obvious sexual dimorphism. Pre-mRNA processing factor 6 (PRPF6) was identified as a coactivator of AR. However, the molecular mechanism underlying the modulation function of PRPF6 on AR-mediated transcriptional activity in HCC needs to be further clarified. In this study, we analyzed data from The Cancer Genome Atlas to show that PRPF6 is highly expressed in HCC. . Our data indicated that PRPF6 interacts with AR/AR splice variants (AR-Vs) and upregulates AR/AR splice variant 7-mediated transcriptional activity even without dihydrotestosterone treatment. We observed that AR is obviously induced by androgen treatment and is mainly expressed in the nucleus in HCC-derived cell lines. Moreover, overexpression of PRPF6 enhances AR expression accompanied with the increase of AR-Vs expression. We provided evidence that PRPF6 participates in upregulating AR self-transcription. PRPF6 facilitates the recruitment of AR to the androgen responsive element region of the AR gene. Finally, PRPF6 depletion inhibits cell proliferation in HCC cells and mouse xenografts. Taken together, our results suggest that PRPF6 as a splicing factor enhances AR self-transcription, thereby coactivating oncogenic AR/AR-Vs actions in HCC.

Although increasing evidence have confirmed that carbon monoxide release molecule-2(CORM-2) plays an active role in the treatment of inflammation and tumors, poor aqueous solubility and short CO-release duration restrict its extensive application. Our previous work synthesized styrene-maleic acid copolymer-encapsulated CORM-2 (SMA/CORM-2) to overcome above-mentioned deficiencies and demonstrated satisfactory effects in colitis. This study is to investigate the function of SMA/CORM-2 on colorectal cancer proliferation and metastasis. CCK-8 experiment is used to clarify the half maximal inhibitory concentration (IC50) of SMA/CORM-2 and to detect cell proliferation. Transwell assay coated with or without matrigel was to detect cell invasion and migration. Western blot was used to detect β-catenin, AKT, p-AKT, VEGF, MMP-2 and MMP-9 proteins. At last, nude mice xenograft was used to further investigate the anti-tumor effect of SMA/CORM-2 in vivo. After SW480 and C26 cells were treated with 0.5 mg/ml SMA/CORM-2, CRC cells proliferation, migration and invasion were inhibited. In vivo, SMA/CORM-2 treatment remarkably suppressed tumor growth and lung metastasis in nude mice. Furthermore, the expression of β-catenin, p-AKT, VEGF, MMP-2 and MMP-9 proteins could be down-regulated after SMA/CORM-2 treatment. SMA/CORM-2 exerted both in vitro and in vivo anti-proliferation and anti-metastatic effects, which may yield a novel therapeutic strategy for CRC.

Objective To obtain the gene expression signatures of NFE2L3 and analyze its clinical significance by mining the data from Oncomine and GEPIA databases. Methods Differential NFE2L3 mRNA expression in colorectal cancer (CRC) was detected based on the Oncomine and GEPIA databases. Correlations between MTERF3 mRNA expression and mismatch repair genes, clinical stages and prognosis in CRC were analyzed by secondary analysis of the GEPIA database. Gene expression differences between cancer and normal tissues were analyzed by the t-test. One-way analysis of variance was used to compare gene expression between different clinical stages. Person correlation analysis was used for correlation study, and Kaplan-Meier method was used for survival analysis. Results A total of 334 different studies were collected in the Oncomine database, of which 55 showed statistically significant expression of NFE2L3. A total of 21 studies involved the expression of NFE2L3 in colorectal cancer tissues and normal tissues, including a total of 1000 samples. Compared with the control group, the expression of NFE2L3 mRNA in colorectal cancer tissues was significantly higher than that in normal colorectal tissues (P 0.05). Conclusion NFE2L3 mRNA is highly expressed in colorectal cancer tissues and is closely related to the expression of mismatch repair genes. Key words: Human nuclear factor erythroid 2-related factor 3; Colorectal cancer; Oncomine; Gene expression profiling, interactive analysis

Purpose B-cell translocation gene 3 (BTG3) has been identified as a candidate driver gene for various cancers, but its specific role in colorectal cancer (CRC) is poorly understood. We aimed to investigate the relationship between expression of BTG3 and clinicopathological features and prognosis, as well as to explore the effects and the role of a possible BTG3 molecular mechanism on aggressive colorectal cancer behavior. Methods BTG3 expression was assessed by immunohistochemistry (IHC) on specimens from 140 patients with CRC. The association of BTG3 expression with clinicopathological features was examined. To confirm the biological role of BTG3 in CRC, two CRC cell lines expressing BTG3 were used and BTG3 expression was knocked down by shRNA. CCK-8, cell cycle, apoptosis, migration, and invasion assays were performed. The influence of BTG3 knockdown was further investigated by genomic microarray to uncover the potential molecular mechanisms underlying BTG3-mediated CRC development and progression. Results BTG3 was downregulated in colorectal cancer tissues and positively correlated with pathological classification ( p = 0.037), depth of invasion ( p = 0.016), distant metastasis ( p = 0.024), TNM stage ( p = 0.007), and overall survival (OS) and disease-free survival (DFS). BTG3 knockdown promoted cell proliferation, migration, invasion, relieved G2 arrest, and inhibited apoptosis in HCT116 and LoVo cells. A genomic microarray analysis showed that numerous tumor-associated signaling pathways and oncogenes were altered by BTG3 knockdown. At the mRNA level, nine genes referred to the extracellular-regulated kinase/mitogen-activated protein kinase pathway were differentially expressed. Western blotting revealed that BTG3 knockdown upregulated PAK2, RPS6KA5, YWHAB, and signal transducer and activator of transcription (STAT)3 protein levels, but downregulated RAP1A, DUSP6, and STAT1 protein expression, which was consistent with the genomic microarray data. Conclusions BTG3 expression might contribute to CRC carcinogenesis. BTG3 knockdown might strengthen the aggressive colorectal cancer behavior.

Objective:To investigate the feasibility,safety and short-term efficacy of hybrid method combined traditional laparoscopy with da Vinci robotic surgical system in rectal neoplasms radical operation.Methods:Retrospectively analysis was made on the clinical data of 268 patients who suffered from rectal neoplasms and underwent da Vinci robotic and laparoscopic radical operation from May 2011 to May 2016.Systematic evaluation was focused on operative procedure,operation selection,complications and short-term recovery.Results:The da Vinci robotic and laparoscopic operation was successful in 267 patients while one was converted to open procedure.The procedure included 199 anterior resections,57 abdominoperineal resections and 12 Hartmann.The mean operative time was (193±33)min,(181±37) min and (173±24) min respectively,and the mean blood loss was (27±8) ml,(36±10) ml and (29±12) ml.The mean number of harvested lymph nodes with or without preoperative neoadjuvant chemoradiotherapy was (11.2±2.8) and (15.4±4.1) respectively.The distal margin was (3.5±0.8) cm without residual cancer cells.Five patients possessed positive circumferential resection margin.Passage of first flatus was (3.0±0.8) d.The time of urinary catheter removed was (3.0±0.6) d.Anastomotic fistula (n=6),intestinal obstruction (n=10) and surgery related infection (n=12) were found without perioperative death.Conclusions:The da Vinci robotic surgical system and laparoscopy is a safe and feasible technique for radical operation of rectal cancer.

Objective To explore compound matrine combined with 5-fluorouracil (5-Fu)on the proliferation and apoptosis of colon cancer cell(SW480) and the expression of CyclinD1.Methods To observe the proliferation and apoptosis of SW480 cell interfered in combined treatment group (compound matrine combined 5-Fu)and the control group (dimethyl sulfoxide combined 5-Fu)using CCK-8cell proliferation experiment and flow cytometry;the expression of CyclinD1 in SW480 cell interfered between the two groups using Western blot.Results The proliferation ability of SW480 interfered in the combined treatment group was lower than that in the control group,the proliferation status on 3 points of 48 hours,72 hours and 96 hours had statistically significant difference between the two groups(P ＜ 0.05).The apoptosis rate of SW480 cells in the combined treatment group(12.7％)was higher than that in the control group(6.18％),the CyclinD1 expression of SW480 interfered in the combined treatment group was less than that in the control group (P ＜ 0.05).Conclusion Compound matrine combined with 5-Fu can significantly inhibit the proliferation of colon cancer cell (SW480) and accelerate the apoptosis of SW480 cell.The mechanism of curative effect on colon cancer may be related to inhibiting the expression of CyclinD1.

Objective To assess the feasibility and possible benefits of a hybrid surgical technique combined traditional laparoscope with robotic surgical system in rectal neoplasms patients. Methods The clinical data of 52 cases underwent hybrid method combined traditional laparoscope with robotic surgical system for rectal neoplasms radical operation from May 2011 to May 2016 in General Hospital of Shenyang Military Area Command were analyzed retrospectively,in which 35 had laparotomy history,11 required splenic flexure mobilization,and 6 needed combined liver resection. Results All the 52 cases underwent operation successfully. The average operative time was(188.6 ± 41.5)min. The average blood loss was(34.3 ± 11.7)mL. The mean numbers of harvested lymph nodes with and without chemoradiation were(11.2 ±2.8)and(15.4 ±4.1). Distal margin and CRM were negative. TNM stage:1 case of stageⅠ, 6 cases of stageⅡ,39 cases of stageⅢ,6 cases of stageⅣ. Passage of first flatus was(2.0±0.5)d. The time to remove a urinary catheter was(3.0 ± 0.6)d. The postoperative hospital stay was(9.0 ± 2.8)d. Except for intestinal obstruction(n=1),no perioperative complications including anastomotic leakage and hemorrhage were present. Conclusion The hybrid approach combined robotic surgical system and laparoscope is a safe and feasible technique for rectal neoplasms radical operation,which may be viable option to patients needing extensive abdominal operation.

美国内战期间,结肠损伤士兵大多数死于感染和败血症.一战期间,结肠损伤的治疗多行Ⅰ期修补,但病死率高达60％～75％.二战期间,结肠损伤多以肠管外置或近端造口作为首选治疗,同时随着抗生素的应用,手术技术、器械的进步以及运送伤员时间的加快,结肠损伤的病死率下降到30％.此后,常规性结肠造口作为治疗结肠损伤的标准术式从未受到质疑.1979年,Stone等[1]发表了一篇有里程碑意义的文章,首次提出结肠损伤的患者行Ⅰ期修补术的疗效优于结肠造口术.但存在以下情况者除外:低血压,出血量＞1L,两个以上器官损伤,严重腹腔污染,手术时间＞8h,损毁性结肠损伤需要切除腹壁大面积缺损.1991年,Chappius等[2]完成了结肠损伤治疗的第2个前瞻性随机研究.结果表明,Ⅰ期修补与结肠造口的并发症发生率无明显的区别.该研究并未将Stone等[1]提出的Ⅰ期修补术禁忌的患者排除在外,并认为没有哪个因素能单独预示Ⅰ期修补术的并发症发生率增加.损伤控制是外科的重要理念,同样被引入到结肠损伤的治疗策略.笔者旨在对结肠损伤的相关问题进行概述.现报道如下.

Objective To explore the expression level of BTG3 protein in colon tumor tissues and its clinical significance. Methods The clinical samples were collected from 63 patients with colon cancer who underwent radical colectomy from 2013 to 2014. The paraffinic specimens of colon cancer which were selected from colon tumor tissues were in the observation group(n=63), and the paraffinic specimens of non tumor which were selected from adjacent nontumorous tissues in the same case as matched control (from the original site 5 centimeters or more)were in the control group(n=63). Using immunohistochemical staining method to com-pare BTG3 protein expression status in the two groups,and to analyze correlation between its expression and the clinical pathological features. Results BTG3 protein was mainly localized in the cytoplasm,the positive expression rate in the observation group(47. 6%) was lower than that in the control group(81. 0%),and the difference was statistically significant(P<0. 01). There was no BTG3 pro-tein positive expression in colonic mucous adenocarcinoma, and the positive expression rate was 52. 6% in colonic carcinoma, the difference was statistically significant(P<0. 05). There was a negative correlation in BTG3 protein expression level,histological type and histological grade of colon cancer(r= -2. 307,P<0. 05;r= -0. 259,P<0. 05). Conclusion BTG3 protein has a lower ex-pression in colon cancer,which is closely related to BTG3 protein expression level,colon cancer tissue histological type and histological grade. BTG3 protein expression level can be used as an auxiliary index of clinical prognosis.

随着微创外科技术的迅速发展，机器人手术逐渐出现在外科各个领域中。目前，以达芬奇为代表的机器人手术系统应用最为广泛。大量临床应用结果证明其具有良好的手术安全性和可靠性。机器人手术系统凭借出色的人体工学设计和先进的技术优势，近10年间在直肠癌的手术应用发展迅猛。笔者就机器人在直肠癌手术中的优缺点、手术方式选择、肿瘤学结局以及术后功能恢复方面，结合自身体会进行探讨。

The daVinci Surgery System is an emerging technology,which has broaden the application of laparoscopy in surgical field.Since FDA approved the daVinci Surgical System in 2000,a number of medical institutions have used it in clinics worldwide and its clinical efficacy and safety have also been approved.Our department has completed 180 robot-assisted operations since the system was introduced into our hospital in 2011.Of these operations,4 were combined resection of organs(2.2%) in which one case was the rectal anterior resection and right hemicolectomy,one was the rectal anterior resection plus splenectomy,one was the rectal anterior re-section and appendectomy and one was the rectal anterior resection and hysterectomy.The most operations of combined resection of or-gans by using traditional laparoscopy were benign tumors.This is because that the radical resection of malignant tumor was relatively dif-ficult and the traditional laparoscopy has limitation to complete such operations.In contrast,the daVinci Surgical System has advantages of 7 degree-of-freedom operation instrument that makes the operation has a great dexterity.Therefore,the system has a great advantage in the operations of combined resection of organs.The operations in the 4 cases were successfully completed.Postoperative recovery was well and no serious complications were found.The authors suggested that the selection of the operation indications still need to be cau-tious and multidisciplinary cooperation and preoperative plan should be noticed.

目的 研究蛋白酶激活受体-2活化剂胰蛋白酶对胃癌MKN28细胞中血管内皮细胞生长因子(VEGF)表达的影响.方法 ①将MKN28细胞分别给予不同浓度(终浓度分别为0、0.1、1.0、10.0及100.0 nmol/L,干预时间均为6h)和不同时间(空白对照组加入PBS溶液,4个干预组胰蛋白酶的终浓度均为10.0 nmol/L,相应的干预时间分别为3、6、12及24 h)的胰蛋白酶干预后,检测MKN28细胞中VEGF mRNA及其蛋白的表达水平,并检测培养液中VEGF蛋白的浓度.②将MKN28细胞分别给予PBS溶液(空白对照组)、胰蛋白酶、胰蛋白酶+PD98059、胰蛋白酶+SB203580、PD98059及SB203580处理,检测MKN28细胞中VEGF mRNA及其蛋白的表达水平.细胞中VEGF mRNA及其蛋白表达水平的检测分别采用荧光定量PCR法和Western blot法,培养液中VEGF蛋白浓度的检测采用酶联免疫吸附实验(ELISA).结果 ①胰蛋白酶浓度的影响.与空白对照组比较,0.1、1.0、10.0及100.0 nmol/L组细胞中VEGF mRNA及其蛋白的表达水平均较高(P＜0.05);与0.1 nmol/L组比较,1.0、10.0及100.0 nmol/L组细胞中VEGF mRNA及其蛋白的表达水平均较高(P＜0.05);与1.0 nmol/L组比较,10.0及100.0 nmol/L组细胞中VEGF mRNA及其蛋白的表达水平均较高(P＜0.05),但10.0 nmol/L组与100.0 nmol/L组细胞中VEGFmRNA及其蛋白的表达水平比较差异均无统计学意义(P＞0.05).空白对照组、0.1 nmol/L组、1.0 nmol/L组、10.0 nmol/L组及100.0 nmol/L组培养液中VEGF蛋白的浓度逐渐递增(P＜0.05).②胰蛋白酶作用时间的影响.空白对照组、3h组、6h组、12h组及24h组细胞中VEGF mRNA的表达水平逐渐递增,5组间两两比较差异均有统计学意义(P＜0.05).与空白对照组比较,3、6、12及24 h组细胞中VEGF蛋白的表达水平均较高(P＜0.05);与3h组比较,6、12及24h组细胞中VEGF蛋白的表达水平均较高(P＜0.05),但6、12及24h组间比较差异无统计学意义(P＞0.05).空白对照组、3h组、6h组、12h组及24h组培养液中VEGF蛋白的浓度逐渐递增,5组间两两比较差异均有统计学意义(P＜0.05).③细胞外调节蛋白激酶(ERK)抑制剂(PD98059)及p38抑制剂(SB203580)的作用.空白对照组、胰蛋白酶+PD98059组、胰蛋白酶+SB203580组、PD98059组和SB203580组细胞中VEGF mRNA及其蛋白的表达水平比较差异均无统计学意义(P＞0.05),但胰蛋白酶组细胞中VEGF mRNA及其蛋白的表达水平均较其余5组相应指标高(P＜0.01).结论 激活MKN28细胞表面的PAR-2可以上调VEGF mRNA及其蛋白的表达,且这一过程受ERK1/2和p38信号通路调控.

机器人外科是一项新兴技术，拓宽了腹腔镜技术在外科领域的应用。达芬奇机器人手术系统自2000年获得FDA 批准后，国外多家医疗机构陆续开展临床应用，其安全性与疗效已得到肯定与共识［1］。沈阳军区总医院自2011年引进该设备，普外科目前共完成达芬奇机器人手术180例，其中腹腔多器官联合切除4例，占2．2％。现将达芬奇机器人手术系统行脏器联合切除4例报道如下。