Effect of Cluster-Zone Leaf Removal at Different Stages on Cabernet Sauvignon and Marselan (Vitis vinifera L.) Grape Phenolic and Volatile Profiles.

This study investigated the effect of leaf removal at three stages of grape development on the phenolic and volatile profiles of Cabernet Sauvignon and Marselan grapevines for two consecutive years in the Jieshi Mountain region, an area of eastern China with high summer rainfall. The results indicated that cluster-zone leaf removal generally reduced the titratable acidity of both varieties, but did not affect the total soluble solids of grape berries. Leaf-removal treatments increased the anthocyanin and flavonol content of berries in both varieties. However, in Cabernet Sauvignon, leaf removal negatively affected the norisoprenoid compounds, with a more pronounced impact observed when the leaf removal was conducted at an early stage. This negative effect may be related to a decrease in the levels of violaxanthin and neoxanthin, potential precursors of vitisprine and ß-damascenone. In contrast, the removal of leaves had no effect on the norisoprenoid aroma of Marselan grapes.

Differences in Aroma Profile of Cabernet Sauvignon Grapes and Wines from Four Plots in Jieshi Mountain Region of Eastern China.

The Bohai Bay region is a famous wine-growing area in China, where the rainfall is concentrated in the summer due to the influence of the temperate semi-humid monsoon climate. As such, the vineyard terrain has a significant impact on the flavor quality of the grapes and the resulting wines. To explore the relationship between the 'Cabernet Sauvignon' wine style and terrain, this study takes four different plots in the Jieshi Mountain region to investigate the differences in the aroma profile of Cabernet Sauvignon grapes and wines of two consecutive vintages. Based on two-way ANOVA, there were 25 free and 8 glycosylated aroma compounds in the grapes and 21 and 10 aroma compounds with an odor activity value greater than 0.1 in the wines at the end of alcohol fermentation (AF) and malolactic fermentation (MLF), respectively, that varied among the four plots. Wines from the four plots showed a significant difference in floral and fruity aroma attributes, which were mainly related to esters with high odor activity values. The difference in concentration of these compounds between plots was more pronounced in 2021 than in 2020, and a similar result was shown on the Shannon-Wiener index, which represents wine aroma diversity. It has been suggested that high rainfall makes the plot effect more pronounced. Pearson's correlation analysis indicated that concentrations of (E)-3-hexen-1-ol in grapes and ethyl 3-methylbutanoate, ethyl hexanoate, isoamyl acetate, isopentanoic acid, and phenethyl acetate in wines were strongly positively correlated with the concentrations of N, P, K, Fe, and electrical conductivity in soil but negatively correlated with soil pH. This study laid a theoretical foundation for further improving the level of vineyard management and grape and wine quality in the Jieshi Mountain region.

PM2.5 aggravates airway inflammation in asthmatic mice: activating NF-κB via MyD88 signaling pathway.

The role of PM2.5 in the bronchial asthma remains unclear. In this study, the deficient mice of TLR4-/-, TLR2-/- and MyD88 -/- were used to establish asthma model. The effects of PM2.5 on the inflammatory response in lung tissue of these mice were observed. PM2.5 increased alveolar macrophages and neutrophils, up-regulated the IL-12 and KC expression in WT mice, but down-regulated their levels in TLR2 -/-, TLR4 -/- and MyD88 -/- mice. OVA+PM2.5 stimulated neutrophil count in WT mice, but it decreased in TLR2 -/- and TLR4 -/- mice. OVA+PM2.5 also increased the Eotaxin, IL-5, IL-13 and MCP-3 expression levels, and OVA specific IgE and IgG1 in serum also increased in WT group. PM2.5 may activate NF-κB through the TLR2/TLR4/MyD88 signaling pathway and aggravate allergic inflammation of lung in asthmatic mice. The microelements in PM2.5 granules, such as lipopolysaccharide, may be an important factor in the high incidence of asthma.

Novel mutation signatures in the prognosis of EGFR-TKIs targeted therapy for non-small cell lung cancer patients based on the 1000-gene panel sequencing.

The application of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer (NSCLC) may be affected by somatic mutations. The purpose of this study was to explore the effect of mutations on the prognosis and tumor markers of NSCLC patients treated with EGFR-TKIs. 21 NSCLC patients treated with EGFR-TKIs were selected, and the targeted sequencing of the tumor tissues or whole blood samples with the 1000-gene panel was conducted to screen mutations. Afterward, functional enrichment analysis was performed based on mutant genes. Subsequently, the correlation between mutations and clinical indicators, prognosis, and tumor markers were analyzed. Finally, the prognosis after taking osimertinib was compared between NSCLC patients with EGFR p.T790M positive and negative mutations, and the EGFR p.T790M concomitant and uncommon mutations were screened. A total of 485 mutations in 251 genes were identified, in which MTOR, AXIN2, AR, EGFR, NOTCH1, and HRAS mutations were significantly correlated with PFS and/or tumor markers. There was no significant difference in PFS, therapeutic effect, and prognosis between EGFR p.T790M positive and negative patients who received osimertinib treatment. Besides, we also found 80 concomitant mutations and 54 uncommon mutations of EGFR p.T790M. AR, HRAS, EGFR, AXIN2, NOTCH1, and MTOR might be key genes to the prognosis of NSCLC treated with EGFR-TKIs. Osimertinib has certain efficacy in EGFR p.T790M negative NSCLC patients.

miR-302a-3p targets FMR1 to regulate pyroptosis of renal tubular epithelial cells induced by hypoxia-reoxygenation injury.

NEW FINDINGS: What is the central question of this study? How does miR-302a-3p play a role in hypoxia-reoxygenation-induced pyroptosis of renal tubular epithelial cells? What is the main finding and its importance? Hypoxia-reoxygenation treatment upregulated the expression of miR-302a-3p in HK-2 cells, and then inhibited the transcription of FMRP translational regulator 1 (FMR1), so as to promote the activation of the NLRP3 inflammasome and aggravate the pyroptosis of HK-2 cells. miR-302a-3p was used as a molecular target in this study, which provides a new theoretical basis for the treatment of renal failure. ABSTRACT: Hypoxia-reoxygenation (H/R) induction can affect miRNA expression and then control NLR family pyrin domain containing 3 (NLRP3) inflammasome-mediated pyroptosis. This study investigated the mechanism of miR-302a-3p in H/R-induced renal tubular epithelial cell (RTEC) pyroptosis. Human HK-2 RTECs were induced by H/R. Lactate dehydrogenase content, cell activity and pyroptosis, and levels of NLRP3, GSDMD-N, caspase-1, interleukin (IL)-1ß, IL-18, superoxide dismutase, and malondialdehyde were detected to verify the effect of H/R on HK-2 cells. The NLRP3 inflammasome action was evaluated after H/R-induced HK-2 cells were treated with BAY11-7082, an inflammasome inhibitor. After inhibiting miR-302a-3p expression, the changes of pyroptosis were observed. The binding relation between miR-302a-3p and FMRP translational regulator 1 (FMR1) was verified. A function-rescue experiment verified the role of FMR1 in the regulation of pyroptosis. H/R-induced HK-2 cells showed significant pyroptosis injury, and the NLRP3 inflammasome was activated. After inhibiting the NLRP3 inflammasome, H/R-induced apoptosis was inhibited. After H/R treatment, miR-302a-3p in HK-2 cells was increased, and miR-302a-3p downregulation limited H/R-induced NLRP3 inflammasome-mediated pyroptosis. FMR1 is the target of miR-302a-3p. Inhibition of FMR1 alleviated the inhibition of H/R-induced HK-2 cell pyroptosis by miR-302a-3p inhibitor. Collectively, inhibiting miR-302a-3p can weaken its targeted inhibition on FMR1, thereby inhibiting the activation of NLRP3 inflammasomes and reducing caspase-1-dependent pyroptosis in HK-2 cells.

Antitumor activity of everolimus in recurrent metastatic endometrial cancer with PTEN deletion: a case report.

Endometrial cancer (EC) is one of the most common gynecological tumors. The first-line treatment for advanced EC is chemoradiotherapy. However, patients in poor health, such as those with intestinal obstruction, have limited tolerance for the side effects of chemoradiotherapy. Individualized precision treatment may bring new hope to these patients. Herein, we have reported on a 56-year-old female patient with metastatic EC combined with severe intestinal obstruction. Due to her inability to tolerate needle biopsy and standard treatment protocols, next-generation sequencing (NGS)-based circulating tumor DNA (ctDNA) testing was performed and PTEN deletion was found. Following, the patient commenced everolimus (10 mg, qd) treatment and partial shrinkage of metastases was observed one month later. Then, everolimus (10 mg, qd) plus carboplatin (100 mg d1, 8, 15, q28d) for 2 cycles, everolimus (10 mg, qd) plus carboplatin (200 mg d1, 8, q21d) for 2 cycles, and everolimus (10 mg, qd) plus carboplatin (200 mg d1, 2, q21d) for 2 cycles were performed, and the patient got partial response for 10 months. From June 2019, the patient continued to benefit from everolimus and subsequently experienced continued benefit for more than 12 months. This is the first reported case of an EC patient who benefited from everolimus as a first-line treatment based on PTEN deletion. This case provides important clinical experience for the precision treatment of patients with advanced EC.

Nivolumab-caused hyperprogression of diffuse large B-cell lymphoma of the testis and spontaneous remission of type 2 diabetes mellitus in an elderly patient: a case report.

Nivolumab has been used in a variety of advanced malignant tumors. Cases of autoimmune diabetes associated with Nivolumab therapy have been reported gradually in recent years. This article reported a case of primary testicular lymphoma in an elderly patient with type 2 diabetes mellitus (T2DM). After treatment with Nivolumab, the primary disease was hyperprogressive disease but the blood glucose was relieved for a long time. Nivolumab may relieve the previous T2DM in diffuse large B-cell lymphoma patients; the potential mechanism needs to be further explored.

Blockade of TRIM59 enhances esophageal cancer cell chemosensitivity to cisplatin by upregulating p53.

Human esophageal cancer (hESC) cell motility adopts various modes, resulting in hESC progression and poor survival. However, how tripartite motif 59 (TRIM59), as the ubiquitination machinery, participates in hESC metastasis is not completely understood. The results indicated that TRIM59 was aberrantly upregulated in hESC tissues compared with adjacent healthy esophageal tissues, which was associated with poor survival and advanced TNM state among patients with hESC. Moreover, patients with hESC with higher TRIM59 expression displayed undetectable p53 expression, which contributed to enhanced progression and motility of hESC. At the molecular level, TRIM59 was indicated to be an E3 putative ubiquitin ligase that targeted the p53 protein, leading to increased degradation of p53, which resulted in decreased chemosensitivity to cisplatin. TRIM59 knockdown reduced TRIM59 expression, increased p53 protein expression, and decreased hESC cell viability, clone formation and migration compared with the small interfering RNA negative control (siNC) group. Furthermore, hESC cell lines were more sensitive to cisplatin in the TRIM59-knockdown group compared with the siNC group. The results indicated a relationship between TRIM59, p53 and the chemosensitivity of cisplatin. The present study suggested that TRIM59 may serve as a promising prognostic indicator for patients with hESC.

KRAS G12C mutations in Asia: a landscape analysis of 11,951 Chinese tumor samples.

BACKGROUND: Kirsten rat sarcoma vial oncogene (KRAS) is one of the most prevalent oncogenes in multiple cancer types, but the incidence is different between the Asian and non-Asian populations. The recent development of KRAS G12C targeting drug has shown great promise. It is thus important to understand the genomic landscape of KRAS G12C in a specific population. METHODS: Sequencing data of 11,951 tumor samples collected from 11/2016 to 7/2019 from multiple centres in China were analyzed for KRAS mutation status. Concomitant genomic aberrations were further analyzed in tumors with KRAS G12C mutations, which were sequenced with comprehensive cancer panel including over 450 cancer-related genes. Smoking status and its correlation with KRAS were analyzed in 2,235 lung cancer cases within this cohort. RESULTS: KRAS mutations were identified in 1978 (16.6%) patient samples. Specifically, KRAS G12C accounted for 14.5% (n=286) of all KRAS mutations. G12C was most commonly seen in lung cancer (4.3%), followed by colorectal cancer (2.5%) and biliary cancer (2.3%). Almost all patients (99.6%) with G12C mutations had concomitant genomic aberrations. These were most commonly associated with the RAS/RTK pathway including BRAF and PI3KCA mutations. Moreover, KRAS mutation was positively correlated with smoking status in lung adenocarcinomas. CONCLUSIONS: The overall incidence of KRAS G12C mutations remains low in the Chinese population. The most common tumor types harboring KRAS G12C mutations are in patients suffering from lung, colorectal and biliary cancers.

[MiR-665 Promotes the Biological Behavior of Small Cell Lung Cancer by Targeting LLGL1].

BACKGROUND: MicroRNAs (miRNAs) are non-coding small molecule RNAs that are widely found in eukaryotic organisms, although some miRNAs have been found in tumors, the expression and effects of miR-665 on small cell lung cancer (SCLC) are unclear. The aim of this study was to analyze the effects of miR-665 on proliferation, cycle, invasion and migration of SCLC cells, and to explore the role of miR-665 in SCLC and its working mechanism. METHODS: The expression of miR-665 in SCLC tissues and adjacent normal tissues was detected by qRT-PCR. TargetScan predicted potential target genes for miR-665 and validated with dual luciferase reporter assays, qRT-PCR and Western blot. CCK8 assay, flow cytometry, Transwell and wound healing assay to detect the effects of miR-665 and LLGL1 on proliferation, invasion, migration and S-phase fraction of SCLC cell line NCI-H446, NCI-H1688. A nude mouse xenograft model of SCLC was constructed and the effect of miR-665 on tumor growth in mice was observed. RESULTS: The expression of miR-665 in SCLC tissues was significantly higher than that in non-tumor normal tissues. MiR-665 could target 3'-UTR of LLGL1 and inhibit its expression. Compared with non-tumor normal tissues, the expression of LLGL1 was significantly lower in SCLC tissues. Inhibition of miR-665 expression could inhibit proliferation, S-phase fraction, invasion and migration ability of SCLC NCL-H446 cells, and interference LLGL1 expression could reverse this inhibition effect. Up-regulation of miR-665 expression could promoted proliferation, S-phase fraction, invasion and migration ability of SCLC NCI-H1688 cells, but this promotion effect was also reversed by overexpression of LLGL1. In a nude mouse xenograft model of SCLC, inhibition of miR-665 expression could up-regulate LLGL1 protein expression and inhibit tumor growth, while up-regulation of miR-665 expression could produce opposite results. CONCLUSIONS: The expression of miR-665 is closely related to SCLC. miR-665 can promote the biological behavior of SCLC cells by inhibiting the expression of target gene LLGL1, and miR-665 play a role in tumor-promoting genes in SCLC.

Survival prediction for patients with lung adenocarcinoma: A prognostic risk model based on gene mutations.

BACKGROUND: Lung adenocarcinoma is the most common type of lung cancer, and it is one of the most aggressive and rapidly fatal tumor types. OBJECTIVE: To identify a signature mutation genes for prognostic prediction of lung adenocarcinoma. METHODS: Four hundred and sixty-two lung adenocarcinoma cases were screened out and downloaded from TCGA database. Mutation data of 18 targeted genes were detected by MuTect. LASSO-COX model was used to screen gene loci, and then a prognosis model was established. Afterwards, 40 clinical patients of lung adenocarcinoma were collected to verify the mutation features and the predictive function of the above prognostic model. The mutations of above 18 genes were sequenced with targeted next generation sequencing (NGS) and analyzed with GATK and MuTect. RESULTS: TP53 (282, 32.38%), NF1 (82, 9.41%) and EGFR (80, 9.18%) were the top 3 most frequent mutation genes. A total of 7 variables were screened out after lasso-COX analysis (tumor stage, age, diagnostic type, SMARCA4, GNAS, PTCH2, TSC2). SMARCA4, GNAS and TSC2 were a gene mutation signature to predict a poor prognosis. CONCLUSIONS: We established a prognostic model for lung adenocarcinoma, and further concluded that SMARCA4, GNAS and TSC2 were a gene signature which plays a prognostic role.

miR-9 regulates the malignant biological behaviors of small cell lung cancer by targeting zinc finger E-box binding homeobox 2 and its possible mechanism / 中国肿瘤生物治疗杂志

@#[Abstract] Objective：To explore the regulatory effect of miR-9 on biological behaviors of small cell lung cancer (SCLC) cells by targeting zinc finger E-box binding homeobox 2 (ZEB2), and to analyze the role of miR-9 in SCLC and its possible mechanism. Methods: qPCR, WB and immunohistochemistry methods were used to detect the mRNA and protein expressions of ZEB2 in cancer tissues and corresponding adjacent tissues of 67 SCLC patients who received surgical treatment at the Department of Oncology, Fourth Hospital of Hebei Medical University from February 2018 to November 2019. TargetScan was used to predict the potential target gene of miR-9, which was later verified by Dual luciferase reporter gene assay, qPCR and WB methods. CCK-8 method, Flow cytometry and Transwell experiment were used to detect the effect of miR-9 and ZEB2 over-expression on the biological behaviors of NCI-H446 cells, and WB was used to detect the protein expressions of E-cadherin, N-cadherin and Vimentin in cells. NCI-H446 cells overexpressing miR-9 were used to construct SCLC nude mouse xenograft model, and the effect of miR-9 on the growth of xenografts was observed. Results: The mRNA and protein expression levels of ZEB2 in SCLC tissues were significantly higher than those in adjacent tissues (P<0.01). There is a potential binding site on the 3' UTR of ZEB2 to bind with miR-9. Compared with the control group, the mRNA and protein expression levels of ZEB2 in NCI-H446 cells of the miR-9 over-expression group were significantly reduced (P<0.01); the proliferation, migration and invasion abilities of NCI-H446 cells were significantly suppressed (P<0.05 or P<0.01), and the expression of EMT protein was reduced; However, simultaneous over-expression of ZEB2 could reverse above effects. In in vivo experiments, the size and weight of transplanted tumors in the miR-9 over-expression group were significantly lower than those in the control group (P<0.05 or P<0.01). The expression of ZEB2 protein in the tumor tissues of nude mice in the miR-9 overexpression group was significantly lower than that in the control group (P<0.01). Conclusion: miR-9 can inhibit the biological behaviors of SCLC cells and the growth of NCI-H446 transplanted tumors in nude mice by targeting and regulating ZEB2.

LncRNA TP73-AS1 interacted with miR-141-3p to promote the proliferation of non-small cell lung cancer.

INTRODUCTION: Recent studies have shown that long non-coding RNAs (lncRNAs) are involved in a variety of biological processes and diseases in humans, including cancer. However, the exact effects and molecular mechanisms of TP73-AS1 in non-small cell lung cancer (NSCLC) progression are still unknown. The present study is aimed to reveal the detailed functions and the mechanism of TP73-AS1 in the regulation of NSCLC cell proliferation. MATERIAL AND METHODS: TP73-AS1 expression in NSCLC tissues and cell lines was determined using real-time PCR assays. The functions of TP73-AS1 in the regulation of NSCLC cell proliferation was evaluated using BrdU assays. The interaction between TP73-AS1 and miR-141-3p was confirmed using luciferase report gene assays. RESULTS: TP73-AS1 was upregulated in NSCLC tissues and cell lines. However, when knockdown of TP73-AS1 inhibited the NSCLC proliferation. By using online tools, we screened out miR-141-3p may combined with TP73-AS1. With use of luciferase assays, we confirmed that miR-141-3p could directly bind to TP73-AS1. In NSCLC tissues, miR-141-3p was down-regulated; TP73-AS1 was inversely correlated with miR-141-3p. CONCLUSIONS: Our data suggest that TP73-AS1 might be an oncogenic lncRNA that promotes proliferation of NSCLC and might be regarded as a therapeutic target in NSCLC.

Novel loss-of-function mutation in BRCA2 gene identified in a Chinese female with a family history of ovarian cancer: A case report.

Inherited loss-of-function mutations in the tumor suppressor BRCA2 gene are associated with a high risk of ovarian cancer in the Chinese population. The current case report discusses a novel heterozygous insertion in BRCA2 gene, c.3195_3196insA, in a 54-year-old Chinese female with hereditary ovarian cancer. This frameshift mutation generates a premature stop codon at amino acid 1,076, which leads to a truncated BRCA2 protein instead of a wild-type BRCA2 protein with 3,418 amino acids. According to the Breast Cancer Information Core database, this mutation has not been previously reported. However, germline mutations of BRCA2 are a more prevalent cause of ovarian cancer in Chinese females compared with females in Western populations. The present study expands the mutational spectra of BRCA2 that is associated with ovarian cancer.

Identification of a novel breast cancer-causing mutation in the BRCA1 gene by targeted next generation sequencing: A case report.

Hereditary breast cancer is an autosomal dominant syndrome caused by germ-line mutations in the human breast cancer genes, BRCA1 and BRCA2. Mutations in either BRCA1 or BRCA2 are the major causes of familial and early-onset breast cancer. The present study investigated a 33-year-old Chinese female patient with breast cancer using targeted next generation sequencing. A novel heterozygous deletion-insertion was also identified in the BRCA1 gene, c.311_312delinsAGGTTTGCA, which causes the formation of a truncated BRCA1 protein of 109 amino acids instead of a wild-type BRCA1 protein of 1,863 amino acids. These results could potentially expand the mutational spectra of BRCA1-associated breast cancer. In addition, these findings may be valuable for the mutation-based screening and genetic diagnosis of breast cancer.

Triple negative breast cancer and immunoglobulin A nephropathy: A case report and literature review.

The association between malignant tumors and the occurrence of glomerular disease has been well documented in previous studies. The most common types of malignant tumor include Hodgkin's lymphoma with minimal change glomerular nephritis, solid tumor with membranous nephropathy and renal cell carcinoma with immunoglobulin (Ig)A nephropathy. The present case study describes a case of a 31-year-old Chinese female patient who was hospitalized with chronic glomerulonephritis. The patient self-administered unknown traditional Chinese medicine; however, protein excretion/24-h remained increased compared with normal levels. After 34 months, a tumor was identified in the patient. Subsequently, the patient was administered breast-conserving surgery and sentinel lymph node biopsy, which validated the diagnosis of triple negative breast cancer at stage IA (T1cN0M0). The patient received chemotherapy and radiotherapy. Following the review of the relevant studies within the last 30 years, it was demonstrated that the present report was the second documented case of breast cancer associated with IgA nephropathy. Thus, the present study hypothesized that IgA nephropathy may be a tumor manifestation in breast cancer.

Expression of miR-133a-3p in gastric cancer tissues and plasma and its effect on proliferation of gastric cancercells / 中国肿瘤生物治疗杂志

@#Objective: To detect the expression of miR-133a-3p in gastric cancer (GC) tissues and plasma of GC patients, and to investigate its effect on the proliferation of GC cells as well as its correlation toprognosis of GC patients. Methods: 52 cases of cancertissues (non-necrosis part) and corresponding adjacent tissues as well as the pre-operative peripheral blood samples from GC patients, who underwent surgery at Department of General Surgery, the Forth Hospital of Hebei Medical University(Shijiazhuang, China) between May 2012 and May 2013, were collected for this study. The plasma sample (n=35) from healthy donors were obtained during their physical examination. RT-qPCR was adopted to detect the expression of miR-133a-3p in gastric cancer tissues, adjacent tissuesand plasma samples of GC patients and healthy volunteers. The relationships between miR-133a-3p expression and the median DFS as well as clinicopathological parameters were also analyzed. CCK-8 assay was adopted to detect the effect of miR-133a-3p silence or over-expression on proliferation of gastric cancer SGC7901 cells. Results: miR-133a-3p was dramatically decreased in gastric cancer tissues (P<0.01), and its expression was associated with TNM stage, tumor infiltration (T), lynphonode metastasis (N), and vascular tumor thrombus (all P<0.01); miR-133a-3p was significantly increased in the plasma of GC patients (P<0.01), and its expression was associated with TNM stage, lynphonode metastasis (N), and vascular tumor thrombus (all P<0.05). miR-133a-3p expression was positively correlated with serum CA199 level of GC patients (P<0.01). The median DFS of patients with high miR-133a-3pexpression in cancer tissues was significantly longer than that of the patients with low expression(20.8 vs 14.8 months, P<0.05); The median DFS of patients with high plasma miR-133a-3p expression was significantly shorter than that of the patients with low expression (14.4 vs 20.3 months, P<0.05). Over-expression of miR-133a-3p could significantly inhibit the proliferation of gastric cancer SGC7901 cells, while miR-133a-3p silence could significantly promote the proliferation (all P<0.05). Conclusion: miR-133a-3p could significantlyinhibit the proliferation of SGC7901 cells; miR-133a-3p aberrantlyexpressed in gastric cancer tissues and plasma, and obviously correlated with prognosis of gastric cancer patients, which may be used as a potential clinical bio-maker for early diagnosis and treatment as well as the prognosis prediction of gastric cancer.

[Sorafenib reverses multidrug resistance of hepatoma cells in vitro].

OBJECTIVE: To explore the role of sorafenib in reversing multidrug resistance (MDR) in hepatoma BEL-7402/FU cells and its possible mechanisms. METHODS: MTT colorimetric assay was used to obtain the dose-response curve of sorafenib in BEL-7402/FU cells, and flow cytometry performed to assess the effect of sorafenib on Rho123 concentration in the cells. The optimal dose of sorafenib for cell treatment was determined according to the results of MTT assay and flow cytometry. MTT assay was employed to evaluate the effect of sorfenib on the cytotoxicity of the antitumor drugs, flow cytometry performed to determine the expression of cell membrane transport protein (P-gp), and RT-PCR used to detect mdr1 gene expression in the cells treated with sorafenib at the optimal dose. RESULTS: Sorafenib at the concentration of 4 micromol/L, efficiently reversed the MDR of the cells with minimal side effects. At the concentration of 4 micromol/L, sorafenib partially reversed the drug resistance of BEL-7402/FU cells to ADM, 5-FU, GEM and DDP, with reversal indexes of 2.98, 7.16, 1.99 and 10.08, respectively. Treatment of the cells with 4 micromol/L, sorafenib also partially down-regulated P-gp expression in BEL-7402/FU cells, and caused a reduction of mdr1 gene expression by 27.3% in comparison with the control cells. CONCLUSION: Sorafenib can reverse MDR in human hepatoma cells probably in association with down-regulation of mdr1 gene expression and increased accumulation of the chemotherapeutic agents in the cells.

[Coadministration of sorafenib and cisplatin inhibits proliferation of hepatocellular carcinoma HepG2 cells in vitro].

OBJECTIVE: To investigate the inhibitory effect of sorafenib in combination with cisplatin (DDP) on the proliferation of hepatocellular carcinoma cells and explore the molecular mechanisms. METHODS: The inhibitory effect of sorafenib and DDP treatment on HepG2 cell proliferation in vitro was assessed by MTT assay. The cell cycle changes and the apoptotic rate of the treated cells were detected by flow cytometry, and the expressions of ERK and pERK examined using Western blotting. RESULTS: Sorafenib and DDP alone both significantly inhibited the proliferation of HepG2 cells, showing a synergistic effect of their actions in combined use (P<0.05). Sorafenib and DDP alone caused cell cycle arrest at G(1) and G(2) phases, respectively. Combined use of the two drugs resulted in significant reduction of the S-phase cell percentage and cell cycle arrest at G(1) and G(2) phases. The coadministration of the drugs significantly increased the apoptosis rate of the cells as compared with the that of the cells with sorafenib or DDP treatment alone (P<0.05). Sorafenib and DDP, used alone or in combination, did not produce obvious effect on ERK expression. Sorafenib treatment for 24 h reduced pERK expression in the HepG2 cells, and the effect was enhanced by combined treatment with sorafenib and DDP. CONCLUSIONS: Sorafenib and DDP show a synergistic effect in inhibiting the proliferation and inducing apoptosis of HepG2 cells. The mechanisms of this synergistic effect can be closely related to the double blockage of the cell cycle and Raf/MEK/ERK pathway inhibition.

[Inhibitory effect of sorafenib combined with arsenic trioxide on hepatocellular carcinoma cells].

OBJECTIVE: To investigate the inhibitory effect of sorafenib in combination with arsenic trioxide (As2O3) on hepatocellular carcinoma cells and explore the mechanisms of the synergetic antitumor effects of the two agents. METHODS: HepG2 cells were treated with sorafenibor, As2O3 alone or their combination, with the untreated cells used as the control. The inhibitory effect of the treatment was analyzed by MTT assay, and the cell apoptosis and mitochondrial transmembrane potential (delta phi m) were detected by flow cytometry. Western blotting was performed to examine the expressions of ERK and pERK in the cells. RESULTS: Sorafenib and As2O3 used alone or in combination both inhibited the proliferation of HepG2 cells, and a synergistic effect of the two agents was noted in their combined action (P<0.05). Combined treatment of the cells resulted in significantly higher apoptsis rate than that in the other groups (P<0.05), and was associated with a more obvious decrease in delta phi m. The expression of ERK was not affected by the two agents used either alone or in combination, but pERK expression was significantly lowered in the cells after combined treatment for 24 h. CONCLUSION: A synergistic effect is observed between the sorafenib and As2O3 in their inhibition of HepG2 cell proliferation, the mechanisms of which may involve reduction of mitochondrial transmembrane potential and Raf/MEK/ERK pathway inhibition.