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1.
Acta Biochim Biophys Sin (Shanghai) ; 56(6): 927-936, 2024 06 25.
Article in English | MEDLINE | ID: mdl-38686459

ABSTRACT

Non-small cell lung cancer (NSCLC) is one of the most prevalent cancers in the world, and early diagnosis can effectively improve patient survival. Here, differentially expressed circIARS genes are screened from the sequencing results, and their molecular characteristics are examined by Sanger sequencing, RNase R assay, agarose gel electrophoresis (AGE), and fluorescence in situ hybridization (FISH). Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) is performed to detect the expression level of circIARS. The diagnostic value of the signature is analyzed using a subject operating characteristic (ROC) curve. Moreover, plasma is collected from postsurgical, chemotherapy, and relapse patients to investigate the prognostic value of circIARS in NSCLC. The expression of circIARS is greater in both the plasma and tissues of NSCLC patients than in those of healthy individuals, and could be used to distinguish NSCLC patients from patients with benign pulmonary disease (BPD), small cell lung cancer (SCLC) patients, and healthy individuals. The expression level of circIARS relatively decreases after antitumor therapy, such as chemotherapy, and relatively increases after recurrence. ROC analysis reveals that circIARS has better detection efficiency than traditional markers. In addition, circIARS expression level is strongly correlated with several clinicopathological parameters. Finally, we tentatively predict the downstream miRNAs or RBP that might bind to circIARS. Plasma circIARS is significantly greater in NSCLC patients and has good stability and specificity as a diagnostic marker, which could aid in the adjuvant diagnosis and dynamic monitoring of NSCLC.


Subject(s)
Biomarkers, Tumor , Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Humans , Carcinoma, Non-Small-Cell Lung/blood , Carcinoma, Non-Small-Cell Lung/diagnosis , Carcinoma, Non-Small-Cell Lung/genetics , Lung Neoplasms/blood , Lung Neoplasms/diagnosis , Lung Neoplasms/genetics , Biomarkers, Tumor/blood , Biomarkers, Tumor/genetics , Female , Male , Middle Aged , Prognosis , Gene Expression Regulation, Neoplastic , RNA, Circular/blood , RNA, Circular/genetics , Aged
2.
J Cancer Res Clin Oncol ; 150(2): 83, 2024 Feb 08.
Article in English | MEDLINE | ID: mdl-38329551

ABSTRACT

BACKGROUND: Circular RNA (circRNA), which has been demonstrated in studies to be abundantly prevalent in tumor cells and bodily fluids and to play a significant role in tumors, has the potential for biological markers to be used to assist tumor diagnosis. This study mainly discusses the potential of circBRIP1 as a biomarker for diagnosing non-small-cell lung cancer (NSCLC). METHODS: First, high-throughput sequencing screened the differentially expressed circBRIP1, and real-time fluorescence quantitative PCR (qRT-PCR) verified its expression in NSCLC. Next, sanger sequencing, agarose gel electrophoresis, RNase R assay, and fluorescence in situ hybridization (FISH) were used to verify its molecular characteristics. The diagnostic value was analyzed by the subject operating characteristic curve (ROC), and the cardinality test was analyzed for correlation with clinicopathological parameters. Finally, we tentatively predicted the downstream miRNA- or RNA-binding protein that may bind to circBRIP1. RESULTS: CircBRIP1 is highly expressed in NSCLC tissues, cells and plasma with good specificity and stability. CircBRIP1 not only can well-distinguish NSCLC patients from benign pulmonary diseases (BPD) patients, healthy individuals and small cell lung cancer (SCLC) patients, but it also has some potential for dynamic monitoring. Combined with the analysis of clinicopathological data, the high level of circRNA expression was related to the degree of tumor differentiation, TNM stage, T stage, lymph node metastasis and distal metastasis in NSCLC patients. In addition, circBRIP1 has a high diagnostic value. CONCLUSIONS: Plasma circBRIP1 is significantly overexpressed in NSCLC patients. It can be used as a sensitive biomarker with unique value for early diagnosis, tumor development and prognosis detection.


Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Humans , Carcinoma, Non-Small-Cell Lung/diagnosis , Carcinoma, Non-Small-Cell Lung/genetics , In Situ Hybridization, Fluorescence , RNA, Circular/genetics , Lung Neoplasms/diagnosis , Lung Neoplasms/genetics , Biomarkers
3.
Anim Genet ; 55(2): 206-216, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38191772

ABSTRACT

Teat number (TNUM) is an important reproductive trait of sows, which affects the weaning survival rate of piglets. In this study, 1166 Dutch Large White pigs with TNUM phenotype were used as the research object. These pigs were genotyped by 50K SNP chip and the chip data were further imputed to the resequencing level. The estimated heritabilities of left teat number (LTN), right teat number (RTN) and total teat number (TTN) were 0.21, 0.19 and 0.3, respectively. Based on chip data, significant SNPs for RTN on SSC2, SSC5, SSC9 and SSC13 were identified using genome-wide association analysis (GWAS). Significant SNPs for TTN were identified on SSC2, SSC5 and SSC7. Based on imputed data, the GWAS identified a significant SNP (rs329158522) for LTN on SSC17, two significant SNPs (rs342855242 and rs80813115) for RTN on SSC2 and SSC9, and two significant SNPs (rs327003548 and rs326943811) for TTN on SSC5 and SSC6. Among them, four novel QTL were discovered. The Bayesian fine-mapping method was used to fine map the QTL identified in the GWAS of the imputed data, and the confidence intervals of QTL affecting LTN (SSC17: 45.22-46.20 Mb), RTN (SSC9: 122.18-122.80 Mb) and TTN (SSC5: 14.01-15.91 Mb, SSC6: 120.06-121.25 Mb) were detected. A total of 52 candidate genes were obtained. Furthermore, we identified five candidate genes, WNT10B, AQP5, FMNL3, NUAK1 and CKAP4, for the first time, which involved in breast development and other related functions by gene annotation. Overall, this study provides new molecular markers for the breeding of teat number in pigs.


Subject(s)
Genome-Wide Association Study , Quantitative Trait Loci , Swine/genetics , Animals , Female , Genome-Wide Association Study/veterinary , Bayes Theorem , Genotype , Phenotype , Polymorphism, Single Nucleotide
4.
J Transl Med ; 21(1): 514, 2023 07 29.
Article in English | MEDLINE | ID: mdl-37516888

ABSTRACT

Programmed cell death (PCD) plays an important role in many aspects of individual development, maintenance of body homeostasis and pathological processes. Ferroptosis is a novel form of PCD characterized by the accumulation of iron-dependent lipid peroxides resulting in lethal cell damage. It contributes to tumor progression in an apoptosis-independent manner. In recent years, an increasing number of non-coding RNAs (ncRNAs) have been demonstrated to mediate the biological process of ferroptosis, hence impacting carcinogenesis, progression, drug resistance, and prognosis. However, the clear regulatory mechanism for this phenomenon remains poorly understood. Moreover, ferroptosis does not usually exist independently. Its interaction with PCD, like apoptosis, necroptosis, autophagy, pyroptosis, and cuproptosis, to destroy cells appears to exist. Furthermore, ncRNA seems to be involved. Here, we review the mechanisms by which ferroptosis occurs, dissect its relationship with other forms of death, summarize the key regulatory roles played by ncRNAs, raise relevant questions and predict possible barriers to its application in the clinic, offering new ideas for targeted tumour therapy.


Subject(s)
Ferroptosis , Neoplasms , Humans , Ferroptosis/genetics , Apoptosis/genetics , Neoplasms/genetics , Carcinogenesis , Pyroptosis , RNA, Untranslated/genetics
5.
BMC Pregnancy Childbirth ; 22(1): 171, 2022 Mar 02.
Article in English | MEDLINE | ID: mdl-35236324

ABSTRACT

OBJECTIVE: The current study was undertaken to investigate the relationship between antimüllerian hormone (AMH) and polycystic ovarian syndrome (PCOS) phenotypes and to determine whether AMH is associated with pregnancy outcomes in infertile women undergoing their first in vitro fertilization (IVF) treatment. METHODS: We performed a retrospective cohort study of 2973 infertile women, including 418 women with PCOS undergoing their first IVF treatment at a private fertility center from January 2014 to March 2018. Women were stratified into three groups using cutoffs defined by the 25th and 75th percentiles of the serum AMH level: 746 women had AMH ≤ 2.25 ng/mL; 1486 women had AMH between 2.25 to 5.71 ng/mL; and 741 women had AMH > 5.71 ng/mL. Endocrine characteristics, PCOS phenotypes, stimulation outcomes, pregnancy outcomes were compared among these groups. When there were any statistical differences (P < 0.05) among the three groups, Bonferroni test was performed as post-hoc tests to determine where the statistical differences existed. To assess the relationships between AMH and pregnancy outcomes in total patients and PCOS patients, logistic regression analysis, adjusted for potential confounding covariates, were performed. RESULTS: Women with high AMH had greater prevalence of hyperandrogenism (HA), polycystic ovarian morphology (PCOM) and amenorrhea than women with low or average AMH. The clinical pregnancy rate were significantly higher in the high-AMH group compared with low- and average-AMH groups (69.9% vs. 58.8% and 64.7% respectively; P < 0.001). The live birth rate was significantly lower in women with AMH ≤ 2.25 ng/mL compared with average- and high-AMH groups (47.6% vs. 55.2 and 59.5% respectively; P < 0.001). However, after controlling for maternal age, oocyte yield, as well as other confounders, AMH was no longer associated with a higher live birth rate (aOR 1.037, 95% CI 0.853-1.261, P = 0.717; aOR 1.099, 95% CI 0.858-1.408, P = 0.455, respectively) and clinical pregnancy rate (aOR 1.064, 95% CI 0.834-1.359, P = 0.617; aOR 1.181, 95% CI 0.875-1.595, P = 0.276, respectively). Moreover, pregnancy outcomes did not differ in PCOS women according to AMH quartiles. CONCLUSION: Increased AMH levels associated with PCOS severity and greater ovarian stimulation. However, AMH was not associated with clinical pregnancy rate and live birth rate after controlling for other confounders in women undergoing IVF. Thus, AMH should not be used to alter clinical decisions and exclude patients based on a low or even undetectable AMH value.


Subject(s)
Anti-Mullerian Hormone/analysis , Anti-Mullerian Hormone/blood , Phenotype , Polycystic Ovary Syndrome/blood , Pregnancy Outcome , Pregnancy Rate , Adult , Embryo Transfer , Female , Fertilization in Vitro/methods , Humans , Infertility, Female/therapy , Pregnancy , Retrospective Studies
6.
Chin Med ; 17(1): 20, 2022 Feb 09.
Article in English | MEDLINE | ID: mdl-35139871

ABSTRACT

BACKGROUND: Buzhongyiqi decoction (BD), Sijunzi decoction (SD), and Shenlingbaizhu decoction (SHD) have been extensively used clinically for the treatment of diseases caused by spleen-Qi deficiency and microbial fermentation has historically been utilized in traditional Chinese medicine (TCM). This study aimed to investigate the mitigative effect of TCM and fermented TCM (FTCM) with Lactobacillus plantarum (LP) on antibiotic-associated diarrhea, and to select an optimal formula and then identify its compounds. METHODS: Dysbacteriosis in mice was induced by ceftriaxone sodium (CS). The mice were then treated with LP, BD, SD, SHD, fermented BD, fermented SD (FSD), and fermented SHD. Diarrhea indexes, the abundances of gut bacteria, intestinal morphometrics, and mRNA expressions of genes related to intestinal barrier function were assessed. Then, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) were employed to identify and relatively quantify the compounds in the selected decoctions. RESULTS: CS significantly increased the fecal output weight, the total number of fecal output, and fecal water content, indicating the occurrence of diarrhea. Bacterial culture tests showed that the above symptoms were accompanied by the disruption of specific intestinal flora. TCM, LP, and FTCM alleviated the diarrhea index and recovered the intestinal microbiota. FTCM showed more advantageous than TCM or LP alone. The mRNA expressions of aquaporins (AQPs) and tight junctions (TJs) decreased by CS were enhanced by TCM, LP, and FTCM. In addition, through UHPLC-Q-TOF/MS, (S)-(-)-2-hydroxyisocaproic acid, L-methionine, 4-guanidinobutyric acid (4GBA), and phenyllactate (PLA) in SD and FSD were identified and relatively quantified. CONCLUSIONS: TCM, LP, and TCM fermented with LP alleviated CS-induced diarrhea symptoms, and improved the intestinal flora and barrier function. Four compounds including (S)-(-)-2-hydroxyisocaproic acid, L-methionine, 4GBA, and PLA in FSD, which were identified by UHPLC-Q-TOF/MS, might function in modulating intestinal flora and improving villi structure.

7.
Huan Jing Ke Xue ; 41(4): 1619-1628, 2020 Apr 08.
Article in Chinese | MEDLINE | ID: mdl-32608667

ABSTRACT

According to the monitoring data of pollutant concentration in the Xiaoqing River estuary from 2008 to 2017, 13 pollutants affecting water quality were selected. The trends of changes in water quality were identified mainly by the pollution index method, and the driving factors affecting water quality changes were quantitatively and qualitatively analyzed by the Spearman and Pearson correlation analysis method and the principal component analysis. The results showed that the water quality of the Xiaoqing River estuary was improved and the pollution was greatly reduced from 2008 to 2017. The pollution factors were still above the limits to varying degrees; COD and petroleum were the most serious pollutants, TP came second, and the remaining pollutants basically met the water quality standards of the Ⅲ. The pollution degree of petroleum, COD, NH4+-N, NO2--N, and As showed an overall downward trend, and the pollution degree of TP, NO3--N, Cu, Pb, Zn, Cd, Hg, and Cr6+ showed an upward trend to different degrees. Social and economic factors were the main driving factors affecting the water quality of the Xiaoqing River. The discharge of pollutants and the changes in land use type led to serious water pollution overall in the Xiaoqing River, whereas the improvement of sewage treatment rate and the implementation of water environmental protection policy were the main factors affecting the water quality improvement of the Xiaoqing River. Natural factors were the secondary driving factors affecting the water quality of the Xiaoqing River. Climate and hydrological factors, such as temperature, sunshine duration, precipitation, and instantaneous runoff, also indirectly affected the water quality to different degrees.

8.
BMC Genomics ; 21(1): 292, 2020 Apr 09.
Article in English | MEDLINE | ID: mdl-32272881

ABSTRACT

BACKGROUND: Improving feed efficiency is one of the important breeding targets for poultry industry. The aim of current study was to investigate the breast muscle transcriptome data of native chickens divergent for feed efficiency. Residual feed intake (RFI) value was calculated for 1008 closely related chickens. The 5 most efficient (LRFI) and 5 least efficient (HRFI) birds were selected for further analysis. Transcriptomic data were generated from breast muscle collected post-slaughter. RESULTS: The differently expressed genes (DEGs) analysis showed that 24 and 325 known genes were significantly up- and down-regulated in LRFI birds. An enrichment analysis of DEGs showed that the genes and pathways related to inflammatory response and immune response were up-regulated in HRFI chickens. Moreover, Gene Set Enrichment Analysis (GSEA) was also employed, which indicated that LRFI chickens increased expression of genes related to mitochondrial function. Furthermore, protein network interaction and function analyses revealed ND2, ND4, CYTB, RAC2, VCAM1, CTSS and TLR4 were key genes for feed efficiency. And the 'phagosome', 'cell adhesion molecules (CAMs)', 'citrate cycle (TCA cycle)' and 'oxidative phosphorylation' were key pathways contributing to the difference in feed efficiency. CONCLUSIONS: In summary, a series of key genes and pathways were identified via bioinformatics analysis. These key genes may influence feed efficiency through deep involvement in ROS production and inflammatory response. Our results suggested that LRFI chickens may synthesize ATP more efficiently and control reactive oxygen species (ROS) production more strictly by enhancing the mitochondrial function in skeletal muscle compared with HRFI chickens. These findings provide some clues for understanding the molecular mechanism of feed efficiency in birds and will be a useful reference data for native chicken breeding.


Subject(s)
Chickens/physiology , Eating , Gene Expression Profiling/veterinary , Gene Regulatory Networks , Animal Feed , Animals , Breeding , Chickens/genetics , Gene Expression Regulation , Muscle, Skeletal/chemistry , Protein Interaction Maps , Sequence Analysis, RNA
9.
J Anim Sci ; 97(9): 3947-3957, 2019 Sep 03.
Article in English | MEDLINE | ID: mdl-31325379

ABSTRACT

Lipid metabolism is considered one of the important factors affecting residual feed intake (RFI). However, the relationship between RFI and expression of lipid metabolism-related genes is unknown in meat-type ducks. To address this issue, a total of 1,000 male meat-type ducks with similar body weight were randomly selected to measure body weight gain and feed intake from 21 to 42 d of age to estimate RFI. The 8 greatest- (high RFI [HRFI]) and lowest- (low RFI [LRFI]) ranking birds were then selected for the present study. Relative expressions of key genes, namely sirtuin 1 (Sirt1), forkhead box O1 (Foxo1), peroxisome proliferator-activated receptor gamma (PPARγ), sterol regulatory element-binding transcription factor 1c (SREBP-1c), fas cell surface death receptor (FAS), acetyl-CoA carboxylase alpha (ACC), carnitine palmitoyltransferase 1A (CPT1A), and acyl-CoA oxidase 1 (ACOX1), were then determined in the HRFI and LRFI ducks by quantitative PCR. The results showed that RFI, feed conversion ratio (FCR), and average daily feed intake (ADFI) were significantly lower (P < 0.05) in LRFI ducks than in HRFI ducks. In addition, expression of Sirt1, Foxo1, CPT1A, and ACOX1 were significantly higher in LRFI ducks than in HRFI ducks (P < 0.05), whereas PPARγ and FAS expression levels were significantly lower in LRFI ducks than in HRFI ducks (P < 0.01). Correlation analysis showed that Sirt1, CPT1A, and ACOX1 expressions were significantly negatively correlated with FCR (r = -0.81 to -0.93; P < 0.01), whereas PPARγ and FAS expressions were significantly positively correlated with FCR (r = 0.74 to 0.87; P < 0.01). PPARγ expression was significantly positively correlated with RFI (r = 0.83; P < 0.01), whereas CPT1A and ACOX1 expressions were significantly negatively correlated with RFI (r = -0.84 to -0.89; P < 0.01). Sirt1 mRNA expression was positively correlated with Foxo1, CPT1A, and ACOX1 mRNA expression (r = 0.78 to 0.92; P < 0.01). Association of Foxo1 with CPT1A and ACOX1 was positive (r = 0.88 to 0.96; P < 0.01). These results suggest that genes related to fatty acid oxidation are upregulated in the liver of ducks with high feed efficiency, while genes associated with lipid synthesis are downregulated. Furthermore, the inclusion of lipid metabolism-related genes in future breeding programs might be beneficial for selecting ducks with greater feed efficiency phenotype.


Subject(s)
Animal Feed/analysis , Ducks/genetics , Eating/genetics , Gene Expression Regulation/genetics , Lipid Metabolism/genetics , Red Meat/analysis , Animals , Body Weight/genetics , Breeding , Ducks/physiology , Fatty Acids/metabolism , Genetic Markers/genetics , Liver/metabolism , Male , Oxidation-Reduction , Phenotype
10.
Anim Sci J ; 90(7): 887-893, 2019 Jul.
Article in English | MEDLINE | ID: mdl-31111649

ABSTRACT

This study was conducted to investigate the effects of free-range system (FRS) and conventional cage-rearing system (CRS) on growth performance, carcass yield, meat quality, lymphoid organ indices, and serum biochemistry of Wannan Yellow chickens. At 56 days of age, a total of 640 male chickens were randomly allocated to FRS and CRS groups, each of which included 4 replicates with 80 chickens in each replicate. The experiment lasted from 56 to 112 days of age. The results showed that CRS chickens exhibited better final body weight, average daily feed intake, average daily gain, and feed conversion ratio, whereas FRS chickens showed better breast and leg yields, shear force, meat color, lower drip loss, and decreased abdominal fat deposition. Moreover, the absolute thymus weight and thymus to body weight ratio of FRS birds were significantly higher than those of CRS birds (p < 0.05). Additionally, FRS chickens had significantly reduced glucose, total protein, triglyceride, and cholesterol contents, but enhanced levels of high-density lipoprotein cholesterol (p < 0.05). In conclusion, the FRS has advantages in breast and leg yields, meat quality, and some serum biochemical parameters of Wannan Yellow chickens, whereas it has negative effects on growth performance.


Subject(s)
Animal Husbandry/methods , Animal Nutritional Physiological Phenomena/physiology , Chickens/growth & development , Chickens/metabolism , Food Quality , Meat , Animal Feed , Animals , Blood Glucose , Blood Proteins , Body Fat Distribution/veterinary , Body Weight , Cholesterol/blood , Eating , Male , Thymus Gland , Triglycerides/blood
11.
Phytochemistry ; 154: 73-76, 2018 Oct.
Article in English | MEDLINE | ID: mdl-30006090

ABSTRACT

Three undescribed aporphine alkaloids laurodionine B (1), illigerine A (2), and N-formyl-laurolitsine (3) were isolated from the methanolic extracts of the Chinese medicinal plant, Illigera aromatica, together with three known analogues (4-6). The chemical structures of 1-6 were identified by spectroscopic methods including 1D and 2D NMR (1H, 13C, COSY, HSQC, and HMBC) and high resolution mass spectrometry (HRESIMS). Compounds 1-3 showed moderate inhibitory activities in vitro against two cultured tumor cell lines, Hela and SMMC7721, with IC50 values of 32.42-62.90 µM. Only compound 1 had in vitro cytotoxic activity against Bcap37 cells, with the IC50 value of 90.61 µM.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Aporphines/pharmacology , Drugs, Chinese Herbal/pharmacology , Hernandiaceae/chemistry , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Aporphines/chemistry , Aporphines/isolation & purification , Cell Proliferation/drug effects , China , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Humans , Medicine, Chinese Traditional , Molecular Structure , Structure-Activity Relationship , Tumor Cells, Cultured
12.
Asian-Australas J Anim Sci ; 31(9): 1401-1406, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29879809

ABSTRACT

OBJECTIVE: The uncoupling protein 3 (UCP3) is a member of the mitochondrial anion carrier superfamily and has crucial effects on growth and feed efficiency in many species. Therefore, the objective of the present study was to examine the association of polymorphisms in the UCP3 gene with feed efficiency in meat-type chickens. METHODS: Six single nucleotide polymorphisms (SNPs) of the UCP3 gene were chosen to be genotyped using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry in meat-type chicken populations with 724 birds in total. Body weight at 49 (BW49) and 70 days of age (BW70) and feed intake (FI) in the interval were collected, then body weight gain (BWG) and feed conversion ratio (FCR) were calculated individually. RESULTS: One SNP with a low minor allele frequency (<1%) was removed by quality control and data filtering. The results showed that rs13997809 of UCP3 was significantly associated with BWG and FCR (p<0.05), and that rs13997811 had significant effects on BW70 and BWG (p<0.05). Rs13997812 of UCP3 was strongly associated with BW70, FI, and FCR (p<0.05). Furthermore, individuals with AA genotype of rs13997809 had significantly higher BWG and lower FCR (p<0.05) than those with AT genotype. The GG individuals showed strongly higher BW70 and BWG than AA birds in rs13997811 (p<0.05). Birds with the TT genotype of rs13997812 had significantly greater BW70 and lower FCR compared with the CT birds (p<0.05). In addition, the TAC haplotype based on rs13997809, rs13997811, and rs13997812 showed significant effects on BW70, FI, and FCR (p<0.05). CONCLUSION: Our results therefore demonstrate important roles for UCP3 polymorphisms in growth and feed efficiency that might be used in meat-type chicken breeding programs.

13.
Oncol Lett ; 15(3): 2821-2828, 2018 Mar.
Article in English | MEDLINE | ID: mdl-29467861

ABSTRACT

The aim of the present study was to investigate the effects of an oleanolic acid derivative, a novel antitumor drug, on the growth of SMMC-7721 human hepatocellular carcinoma cells and the underlying mechanism. An MTT assay was performed to determine the cytotoxicity of the oleanolic acid derivative. Cell membrane integrity was assessed using fluorescence microscopy to assess the uptake of annexin V-FITC/propidium iodide (PI). Western blotting was used to detect the apoptosis-associated proteins B cell lymphoma-2 (Bcl-2), Bax, caspase-9 and caspase-3. A spectrophotometer was used to analyze the intracellular adenosine triphosphate (ATP) expression level. The loss of mitochondrial membrane potential was detected by performing the JC-1 assay. ELISA was used to evaluate the content of cytochrome c (Cyt-C). The oleanolic acid derivative reduced the cell viability of SMMC-7721 cells in a dose- and time-dependent manner. The half maximal inhibitory concentration values of the oleanolic acid derivative in SMMC-7721 cells at 24, 48 and 72 h were 26.80, 11.85, and 6.66 µM, respectively. The antiapoptotic-protein Bcl-2 was downregulated, and the proapoptotic protein Bax was upregulated following treatment with the oleanolic acid derivative for 48 h. The oleanolic acid derivative induced the cleavage of caspase-9 and caspase-3 as well as promoted annexin V-FITC/PI uptake in SMMC-7721 cells. Furthermore, treatment of SMMC-7721 cells with the oleanolic acid derivative induced a reduction of the intracellular ATP expression level, loss of ΔΨm and Cyt-C release from the mitochondria. The oleanolic acid derivative induced apoptosis in SMMC-7721 human cells. Mitochondrial dysfunction was involved in the anticancer effects of this derivative on SMMC-7721 human cells.

14.
Oncol Lett ; 14(2): 1731-1736, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28789402

ABSTRACT

The aims of the present study were to investigate the effects of esculetin on the proliferation of human hepatocellular carcinoma SMMC-7721 cells and to determine the underlying mechanism behind this activity. An MTT assay was used to assess cell proliferation, reverse transcription-quantitative polymerase chain reaction was used to determine the relative mRNA expression levels of ß-catenin, c-Myc and cyclin D1, and western blot analysis was utilized to determine the levels of the associated proteins. Compared with the dimethyl sulfoxide control, esculetin reduced the cell viability of SMMC-7721 and HL-7702 cells in a dose- and time-dependent manner. Treatment of SMMC-7721 cells with esculetin resulted in downregulation of the mRNA and protein levels of ß-catenin, c-Myc and cyclin D1. Esculetin increased the phosphorylation of ß-catenin at Ser33/Ser37/Thr41 and inhibited the proliferation of human hepatoma SMMC-7721 cells by suppressing the Wnt signaling pathway. The results of the present study suggest that esculetin inhibited the Wnt/ß-catenin signaling pathway in SMMC-7721 cells and may have potential as an effective anti-cancer drug, acting to inhibit the Wnt/ß-catenin signaling pathway.

15.
Pharmazie ; 71(7): 398-401, 2016 Jul 07.
Article in English | MEDLINE | ID: mdl-29441916

ABSTRACT

Oleanolic acid, isolated from privet, has shown antitumor effects in several cancers. However, the underlying molecular mechanism associated with these effects is largely unknown. In this study, we explored the effect of oleanolic acid derivatives on the Wnt/ß-catenin signaling pathway in human hepatocellular carcinoma SMMC-7721 cells. The mRNA and protein levels of related genes were determined by real-time quantitative PCR and Western blot, respectively. Treatment of SMMC-7721 cells with oleanolic acid derivatives led to the downregulation of the mRNA and protein levels of ß-catenin, c-myc, and cyclin D1. Treatment with oleanolic acid derivatives decreased the levels of ß-catenin in both the cytoplasm and the nucleus. Moreover, oleanolic acid derivatives promoted the phosphorylation of ß-catenin (Ser33/37/Thr41) in the cytoplasm. Our results suggest that oleanolic acid derivatives inhibit the Wnt/ß-catenin signaling pathway by stimulating the phosphorylation of ß-catenin (Ser33/37/Thr41) in human SMMC-7721 cells.


Subject(s)
Oleanolic Acid/analogs & derivatives , Oleanolic Acid/pharmacology , Wnt Signaling Pathway/drug effects , beta Catenin/drug effects , beta Catenin/metabolism , Cell Line, Tumor , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Cyclin D1/antagonists & inhibitors , Cyclin D1/biosynthesis , Cytoplasm/drug effects , Cytoplasm/metabolism , Down-Regulation/drug effects , Gene Expression Regulation, Neoplastic , Humans , Phosphorylation/drug effects , Proto-Oncogene Proteins c-myc/antagonists & inhibitors , Proto-Oncogene Proteins c-myc/biosynthesis , RNA, Messenger/biosynthesis , RNA, Messenger/genetics
16.
Cell Commun Adhes ; 19(5-6): 79-84, 2012 Oct.
Article in English | MEDLINE | ID: mdl-23150925

ABSTRACT

The immune adherence (IA) between the porcine erythrocytes and the opsonized Escherichia coli carried green fluorescent protein gene (GFP-E.coli) were detected by the fluorescence microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) with an attempt to verify the existence of IA between the porcine erythrocytes and complemented-opsonized microbes. Under fluorescence microscopy, GFP-E.coli opsonized by fresh rabbit serum complement adhered to the erythrocytes and could not be detached by PBS washing, and no IA was observed between the erythrocytes and nonopsonized GFP-E.coli after co-incubation. SEM and TEM also revealed the existence of IA between the serum complement-opsonized GFP-E.coli membrane and the erythrocyte membrane. The partial complement receptor type 1 (CR1)-like gene from porcine was generated by RT-PCR and rapid amplification of cDNA 3' end (3' RACE) (157bp and 578bp), both of which have high similarity with published mammal's CR1 gene. The sequences were spliced based on homology comparison and submitted to GenBank (GenBank Accession No. JX033989). These results indicated that the porcine erythrocytes were able to bind to the opsonized microorganisms. Furthermore, the sequencing results confirmed that the CR1-like gene exists in porcine.


Subject(s)
Erythrocytes/immunology , Immune Adherence Reaction , Animals , Base Sequence , Cell Adhesion , Complement System Proteins/immunology , Complement System Proteins/metabolism , Erythrocyte Membrane/immunology , Erythrocytes/cytology , Erythrocytes/metabolism , Escherichia coli/immunology , Escherichia coli/metabolism , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Humans , Microscopy, Fluorescence , Molecular Sequence Data , Opsonin Proteins/metabolism , Rabbits , Receptors, Complement/genetics , Receptors, Complement/metabolism , Sequence Homology , Swine , Tribolium/genetics
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