Correlation between pre-treatment serum total blood bilirubin and unconjugated bilirubin and prognosis in patients with colorectal cancer.

BACKGROUND: Epidemiological studies have found that unconjugated bilirubin (UCB) levels are positively correlated with the incidence of colorectal cancer (CRC). Therefore, bilirubin may also play an important role in the prognosis of CRC. AIM: To investigate the predictive value of total bilirubin (TBIL) and UCB in the prognosis of patients with CRC. METHODS: A total of 142 CRC patients were selected as the research subjects in Jingxian Hospital, from October 2014 to May 2021. General and tumour-related clinical data at admission and the overall survival at 3 years after surgery were collected. The optimal cut-off values of TBIL and UCB were determined by receiver operating characteristic curve analysis. Univariate and multivariate Cox regression were used to analyse the effect of bilirubin level on the survival of CRC patients. The Kaplan-Meier method was used to assess the survival time. RESULTS: The 3-year overall survival rate of CRC patients was significantly higher in the high TBIL (> 13.45 µmol/L) group than in the low TBIL (≤ 13.45 µmol/L) group (76.4% vs 37.1%; P < 0.05). The 3-year overall survival rate of CRC patients in the high UCB (> 10.75 µmol/L) group was significantly higher than that in the low UCB (≤ 10.75 µmol/L) group (83.3% vs 34.2%; P < 0.05). Multivariate Cox regression analysis showed that higher TBIL levels were an independent predictor of better prognosis in CRC patients (hazard ratio = 0.360, 95% confidence interval: 0.159-0.812, P = 0.014). CONCLUSION: TBIL levels can be used as a prognostic indicator for CRC patients.

Secreted frizzled-related protein 1 (SFRP1) gene methylation changes in the human lung adenocarcinoma cells treated with L-securinine.

Lung cancer remains the leading cause of cancer-related death worldwide. It is important to explore the biomarkers of diagnosis and prognosis in lung cancer. To evaluate the cytotoxicity of L-securinine and the expression and methylation of secreted frizzled-related proteins (SFRPs) genes in the human lung adenocarcinoma cells, cell counting kit-8 (CCK-8) assay was used to assess the proliferation of lung adenocarcinoma cells treated with L-securinine. Quantitative real-time PCR (qRT-PCR) and bisulfite sequencing PCR were used to detect the expression and the DNA methylation of SFRPs genes, respectively. L-securinine inhibited the proliferation of lung adenocarcinoma cells and induced the upregulation of SFRP1 gene expression and the methylation changes at CpG sites in the SFRP1 promoter region. L-securinine was a potential agent in the treatment of lung cancer by upregulation of SFRP1 gene expression and changing the SFRP1 gene methylation.

Gracillin induces apoptosis in HL60 human leukemic cell line via oxidative stress and cell cycle arrest of G1.

Gracillin, a kind of steroidal saponin isolated from the root bark of wild yam Dioscorea nipponica has been reported to exert antitumor activity. In the present study, we investigated the anticancer activity of gracillin against HL60 cells, and evaluated the possible mechanism involved in its antineoplastic action. The cell proliferation was evaluated by cell counting Kit-8 (CCK-8) assay, gracillin inhibited the growth of HL60 cells in a time- and concentration-dependent manner. Flow cytometry was used to analyze the cell cycle distribution whereas Annexin V-FITC/PI flow cytometry analysis was carried out to confirm apoptosis induced by gracillin, Our results demonstrated that gracillin could induce cell cycle arrest of G1 and apoptosis in HL60 cells. Furthermore, based on the biochemical methods, induction of oxidative stress by gracillin was indicated by increased the content of malondialdehyde (MDA), and decreased superoxide dismutase (SOD) activity. In addition, real time-PCR verified the expression of apoptosis-related genes, the mRNA level of Bcl-2 was decreased dramatically, while Bax was remarkably increased by gracillin. Taken together, gracillin could induce cell cycle arrest, oxidative stress, and apoptosis in HL60 cells, and has the potential to be developed as an antitumor agent.

Effects of intracavitary administration of Endostar combined with cisplatin in malignant pleural effusion and ascites.

This study evaluated the efficacy and safety of intracavitary administration of recombinant human endostatin (Endostar) combined with cisplatin chemotherapy in treating malignant pleural effusion and ascites. Forty-five patients with malignant pleural effusion and ascites were divided into the EP group (n = 23), who received Endostar and cisplatin intracavitarily, and P group (n = 22), who were intracavitarily treated with cisplatin only. Pleural effusion and ascites were completely drained before treatments. The treatment was administered once a week; two treatments were considered as one course. The outcome quality of life as well as toxicity were evaluated. The objective overall response and disease control rates were, respectively, 78.3 % (18/23) and 87.0 % (20/23) in EP group. In contrast, these parameters were significantly (p < 0.05) lower in P groups: 40.9 % (9/22) and 59.1 % (13/22), respectively. The improvement rate of Karnofsky Performance Status was 87.0 % (20/23) in EP group versus 59.1 % (13/22) in P group (p < 0.05). All patients tolerated the combined treatment well, and no severe adverse effects were observed. Intracavitary injection of Endostar combined with cisplatin is effective and safe to treat malignant pleural effusion and ascites.

Virosecurinine induces apoptosis by affecting Bcl-2 and Bax expression in human colon cancer SW480 cells.

Virosecurinine, the major alkaloid isolated from Securinega suffruticosa Pall Rehd was found to exhibit growth inhibition and cytotoxicity against huaman colon cancer SW480 cells via the microculture tetrazolium (MTT) assay. Due to its greater cytotoxic potency and selectivity towards SW480 cells, flow cytometry was used to analyze the cell cycle distribution of control and treated SW480 cells whereas Annexin V-FITC/PI flow cytometry analysis was carried out to confirm apoptosis induced by virosecurinine in SW480 cells. Apoptotic regulatory genes were determined by RT-PCR analysis. Virosecurinine was found to induce G1/S cell cycle arrest which led to predominantly apoptotic mode of cell death. Mechanistically, virosecurinine was found to up-regulated the Bax gene expression and down-regulated the Bcl-2 expression in SW480, The ratio of Bcl-2 to Bax was significantly decreased. Hence, we suggest that virosecurinine induced apoptosis in SW480 cells by affecting the expression of bcl-2 and bax.

23-hydroxybetulinic acid-induced HL-60 cell autophagic apoptosis and its molecular mechanism.

This study investigates the effects of 23-hydroxybetulinic acid (23-HBA) in inducing HL-60 autophagic apoptosis and explores its potential molecular mechanism. The generation depression effects of HL-60 cells cultured in vitro were detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method (p < 0.05), and dosage time-dependent relationships were found. Numerous autophagic vacuoles and empty vacuoles were observed in HL-60 cells treated with 12.5 µM 23-HBA for 24 h by electron microscopy; it was also observed that the process of cell division diminished. Using flow cytometry, the number of apoptotic cells was seen to obviously increase, and the G1/S phase was retarded. It was observed that 23-HBA tended to arrest cells at the G1 phase of the cell cycle. The percentage of the apoptotic cells increased as treatment duration and concentration increased. Beclin-1 expression enhanced with an increase in 23-HBA concentration. We can conclude that 23-HBA can induce autophagic apoptosis in HL-60; the autophagic apoptosis induced by 23-HBA is related to the upregulation of expression of the apoptotic gene beclin-1.

L-securinine induced the human colon cancer SW480 cell autophagy and its molecular mechanism.

AIMS: To investigate the anti-tumor effects of L-securinine inducing colon cancer SW480 cell autophagy and explore its potential molecular mechanism. MAIN METHODS: MTT method was used to detect the antitumor effect of SW480 cells cultured with L-securinine in vitro. Light and electron microscopy were used to observe SW480 cells treated with L-securinine morphological changes. Flow cytometry was used to observe the apoptoticratio and cell cycle inducing with the L-securinine in SW480 cells, and the autophagic apoptosis ratio was determined by FITC-conjugated annexin V by flow cytometry (FCM). FCM was applied to analysis cell cycle; the expression of autophagy gene Beclin-1 was examined by reverse transcriptase polymerase chain reaction (RT-PCR). KEY FINDINGS: The generation depression effects of SW480 cells cultured in vitro were detected byMTT method (Pb0.05), and there were dosage-time dependent relationships. Numerous autophagic vacuoles and empty vacuoles were observed in SW480 cells treated with 2.5 µM L-securinine for 48 h by electron microscopy, and the process of cell division that got less was observed.Through flow cytometry, a number of observed autophagic cells were obviously increased, and G1/S phase was retarded. L-Securinine tended to arrest cells at the G1 phase of the cell cycle. The percentage of the apoptotic cells increased as treatment duration and concentrations increased. Beclin-1 expression enhanced with L-securinine concentration increased. SIGNIFICANCE: L-Securinine has an anti-tumor effect against colon cancer SW480 cell. The L-securinine can induce striking autophagy in SW480 cell in vitro. The autophagy induced by L-securinine is related with upregulating the expression of autophagy gene Beclin-1.

In vitro estrogenic activity of formononetin by two bioassay systems.

AIM: To study the estrogenic activity of formononetin in vitro. METHODS: We have established a highly sensitive bioassay system by placing estrogen-responsive elements upstream of the luciferase reporter gene, and used this assay to determine the estrogenic activity of formononetin. Cell growth was measured by the MTT (3-(4,5-dimethylthioazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and MG-63 cell function was studied by measuring alkaline phosphatase activity. RESULTS: Formononetin activated expression of the estrogen-responsive reporter gene in human breast cell line MCF-7 in a concentration-dependent manner (0.5-500 microM), and this activation was inhibited by estrogen antagonist (ICI 182780 at 100 nM). Furthermore, it induced the proliferation of MCF-7 breast cancer cells and MG-63 osteosarcoma cells, and it also increased the alkaline phosphatase activity in MG-63 cells. CONCLUSION: Formononetin is a phytoestrogen that exhibits variable degrees of estrogen receptor agonism in different test systems.

Up-regulation of P-glycoprotein expression by glutathione depletion-induced oxidative stress in rat brain microvessel endothelial cells.

Glutathione (GSH) depletion has been implicated in the pathogenesis of neurological diseases. During GSH depletion, cells of the blood-brain barrier (BBB) are subjected to chronic oxidative stress. In this study, we investigated the effect of such stress, produced with the GSH synthesis inhibitor l-buthionine-(S,R)-sulfoximine (BSO), on expression of P-glycoprotein (Pgp) in primary cultured rat brain microvessel endothelial cells that comprise the blood-brain barrier (BBB). Application of BSO to cell monolayers at concentrations up to 800 microm caused increases in expression of Pgp. Concentrations >or= 400 microm BSO decreased cell viability. Application of 200 microm BSO caused a significant increase in Pgp function activity, as assessed by rhodamine 123 (Rh123) accumulation experiments. At this concentration, BSO produced time-dependent decreases in levels of intracellular GSH and increases in levels of intracellular reactive oxygen species (iROS). The increases were also observed within 48 h following BSO treatment in mdr1a and mdr1b mRNA. Exposure of cells to BSO for 24 h produced maximal effects in the accumulation of iROS, and in expression and function of Pgp. The ROS scavenger N-acetylcysteine prevented ROS generation and attenuated the changes of both expression and activity of Pgp induced by BSO. Therefore, the transport of Pgp substrates may be affected by changing Pgp expression under conditions of chronic oxidative stress induced by GSH depletion.

Patensin-induced apoptosis is accompanied by decreased bcl-2 expression and telomerase activity in HL-60 cells.

The present study aimed to investigate the effect of telomerase activity in patensin-induced apoptosis and the regulation of B cell leukemia/lymphoma 2 (bcl-2) gene expression in human leukemia HL-60 cells. Apoptosis of HL-60 cells was induced by patensin (100 micromol L(-1)) for 3, 6, 12 and 24 h. Apoptosis and bcl-2 were determined by flow cytometry analysis. A polymerase-chain-reaction-based telomeric repeat amplification protocol assay was used to detect the telomerase activity. Patensin induced growth arrest and apoptotic cell death in HL-60 cells. The telomerase activity was inhibited in a time-dependent manner during the patensin-induced apoptosis of HL-60 cells, and the expression of bcl-2 was progressively down-regulated by patensin. Inhibition of the telomerase activity of HL-60 cells was closely related to the patensin-induced apoptosis. The present results indicate that inhibition in telomerase and reduced bcl-2 gene expression may play a role in the patensin-induced apoptosis of HL-60 cells.

23-Hydroxybetulinic acid-mediated apoptosis is accompanied by decreases in bcl-2 expression and telomerase activity in HL-60 Cells.

23-Hydroxybetulinic acid, a derivative of betulinic acid, was investigated for its apoptotic effect and the associated telomerase activity in human leukemia HL-60 cells. Apoptosis and bcl-2 were determined by flow cytometry analysis. A PCR-based telomeric repeat amplification protocol assay was used to detect telomerase activity. Results showed that 23-hydroxybetulinic acid induced growth arrest and apoptotic cell death in HL-60 cells. The apoptotic events were associated with concurrent down-regulation of bcl-2 and the telomerase activity. Our data suggest that 23-hydroxybetulinic acid may be a potential cytotoxic agent for treatment of cancer.

Daidzin protects PC12 cells from serum deprivation-induced apoptosis.

This article examines the effect of daidzin on PC12 cell apoptosis induced by serum-free medium. PC12 cell survival was measured by MTT assay. The DNA content and percentage of apoptosis were monitored by flow cytometry and DNA fragmentation was analyzed by agarose gel electrophoresis. The results showed that serum-free (12 h) medium induced apoptosis in PC12 cells. When the cells had been treated with daidzin (0.1, 1 microM) for 12 h, the percentage of PC12 cell apoptosis was significantly decreased to 12.21 and 4.24% from 91.94% in the group with serum deprivation, and DNA fragmentation was prevented. Daidzin (0.01-10 microM) attenuated the cytotoxic effect of sodium cyanide (20 mM), glutamate (0.5 mM) and sodium nitroprusside (0.5 mM) in a manner dependent on concentration. The results suggested that daidzin prevented PC12 cell from serum free-induced apoptosis.

Inhibition of telomerase activity and bcl-2 expression in berbamine-induced apoptosis in HL-60 cells.

The present study was aimed to investigate the effect of telomerase activity in berbamine-induced apoptosis and the regulation of B cell leukemia/lymphoma 2 ( bcl-2) gene expression in human leukemia HL-60 cells. Apoptosis of HL-60 cells was induced by berbamine (10 microM) for 3, 6, 12 and 24 h. Apoptosis and bcl-2 were determined by flow cytometry analysis. A polymerase chain reaction-based telomeric repeat amplification protocol assay was used to detect the telomerase activity. Berbamine induced growth arrest and apoptotic cell death in HL-60 cells. The telomerase activity was inhibited in a time-dependent manner during the berbamine-induced apoptosis of HL-60 cells, and the expression of bcl-2 was progressively down-regulated by berbamine. Inhibition of the telomerase activity of HL-60 cells was closely related to the berbamine-induced apoptosis. The present results indicate that inhibition of telomerase and reduced bcl-2 gene expression may play a role in the berbamine-induced apoptosis of HL-60 cells.