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1.
Article in Chinese | WPRIM | ID: wpr-1031485

ABSTRACT

ObjectiveTo explore the possible mechanism of Shipi Xiaoji Beverage (实脾消积饮) in treating primary liver cancer from the perspective of mitochondrial dynamics imbalance and ferroptosis. MethodsThe Shipi Xiaoji Beverage-containing serum was prepared, and liver cancer HepG2 cells were cultured in vitro. The blank group, control group, cisplatin group (10 µg/ml), and 5%, 10%, and 15% medicated serum groups were set, with 4 duplicate holes in each group. After adding the corresponding medicinals to each group, the cell survival rate was calculated using the CCK-8 test after 24-hour culture at 37 ℃ with 5% CO2. Using transwell invasion assay, the number of cells that reached the upper chamber membrane through penetrating the Matrigel was calculated after culturing for 48 hours. The scratch migration rate was calculated after incubation for 24 h and 48 h in the scratch experiment, and the concentration of the medicated serum was screened for subsequent experiments. The control group, 10% medicated serum group (the screened concentration) and 10% medicated serum+Mdivi-1 group were set up. After culturing for 24 hours, the average fluorescence intensity of mitochondria and reactive oxygen species (ROS), adenosine triphosphate (ATP) content and level were measured, and the protein expression of cytoplasmic and mitochondrial dynein-related protein 1 (Drp1) and mitochondrial phosphorylated dynamin-related protein 1 (p-Drp1) was detected. The control group, 10% medicated serum group, ferroptosis activator group (Erastin, 10 μmol/L) and 10% medicated serum+ferroptosis inhibitor group (fer-1,10 μmol/L) were set up, and after adding the corresponding medicinals to each group and culture for 24 hours, the intracellular glutathione (GSH), malondialdehyde (MDA) content and ferrous ion level were detected, as well as the protein expression of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) in cells. ResultsCompared to those in the control group, the cell survival rate, invasion number, 24 h and 48 h scratch migration rate in all medicated serum groups and cisplatin group significantly decreased (P<0.01); as the concentration of medicated serum increased, the cell survival rate, invasion number and 48h scratch migration rate of each medicated serum group gradually decreased (P<0.01), and finally, 10% of Shipi Xiaoji Beverage-containing serum was selected for subsequent experimental concentration. Compared to the control group, the 10% medicated serum group had decreased average fluorescence intensity of mitochondria and ATP content and increased average fluorescence intensity of ROS (P<0.05); compared to the 10% medicated serum group, the 10% medicated serum+Mdivi-1 group had higher average fluorescence intensity of mitochondria and ATP content and lower average fluorescence intensity of ROS (P<0.05 or P<0.01). Compared to those in the control group, GSH content and protein expression of SLC7A11 and GPX4 in other groups significantly decreased, while MDA content and ferrous ion level increased (P<0.05 or P<0.01); compared to the ferroptosis activator group, the 10% medicated serum group and the 10% medicated serum+ferroptosis inhibitor group had decreased MDA content and ferrous ion level, and increased protein expression of SLC7A11 and GPX4 (P<0.05 or P<0.01); the 10% medicated serum+ferroptosis inhibitor group had higher GSH level than the 10% medicated serum group (P<0.01). ConclusionShipi Xiaoji Beverage may inhibit the proliferation, infestation, migration of HepG2 cells by causing mitochondrial dynamics imbalance and inducing ferroptosis, thereby playing against liver cancer.

2.
Article in Chinese | WPRIM | ID: wpr-1017335

ABSTRACT

Objective:To discuss the factors related to the prognosis in the alpha fetoprotein(AFP)negative hepatocellular carcinoma(HCC)patients,and to construct the nomogram for predicting the survival time of the patients.Methods:The retrospective analysis on data of 2 064 cases of AFP negative HCC patients extracted from the Surveillance,Epidemiology,and End Results(SEER)Database was conducted,and all the patients were divided into training cohort and internal validation cohort at a ratio of 7∶3,and 101 AFP negative HCC patients from the Integrated Traditional Chinese and Western Medicine Hospital in Hunan Province were regarded as the external validation cohort.The univariate Cox regression analysis results were incorporated into the multivariate analysis,and the independent risk factors for the AFP negative HCC patients were obtained by multivariate Cox analysis to build a cancer specific survival(CSS)prognosis nomogram for the AFP negative HCC patients.The predictive efficacy and clinical utility of the nomogram were evaluated by time-dependent receiver operating characteristic curve(ROC),calibration plots,and decision curve analysis(DCA).The total score obtained from the nomogram was used for the risk stratification to compare the degree of risk discrimination between the nomogram and the American Joint Committee on Cancer(AJCC)staging system.Results:Ten independent risk factors were selected by multivariate Cox regression analysis to construct 3-year,4-year,and 5-year CSS prognostic nomograms for the AFP negative HCC patients,including the patient's age,pathological grade,surgical status,radiotherapy status,chemotherapy status,lung metastasis status,tumor size,tumor T stage,tumor M stage,and marital status.The area under curve(AUC)for the 3-year,4-year,and 5-year time-dependent ROC in the training cohort were 0.807(95%CI:0.786-0.828),0.804(95%CI:0.782-0.826),and 0.813(95%CI:0.790-0.835),respectively.In the internal validation cohort,they were 0.776(95%CI:0.743-0.810),0.772(95%CI:0.737-0.808),and 0.789(95%CI:0.752-0.826),and in the external validation cohort,they were 0.773(95%CI:0.677-0.868),0.746(95%CI:0.620-0.872),and 0.736(95%CI:0.577-0.895).The calibration plots verified that the nomogram fitted well with the perfect line.The DCA curve revealed that the net benefit of the nomogram was significatly higer than that of the AJCC staging system at certain probability thresholds compared with AJCC staging,the nomogram had a better ability to identify high-risk individuals.Conclusion:The serum AFP expression is one of the prognostic markers for the HCC patients.For those patients with AFP negative expression in serum,different considerations should be taken.The nomogram model based on multiple risk factors is a promising clinical tool for assessing the CSS in the AFP negative HCC patients.

3.
Article in Chinese | WPRIM | ID: wpr-1023875

ABSTRACT

AIM:To investigate the mechanism through which esculetin induces ferroptosis of mouse breast cancer 4T1 cells.METHODS:Molecular docking of esculetin with p53,solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)proteins was performed,and Kaplan-Meier curves and time-dependent receiver oper-ating characteristic curves were drawn.The 4T1 cells were divided into 6 groups,namely the control(CON),1/2 IC50,IC50,2 IC50,erastin,and capecitabine groups.The appropriate drug concentration for treating 4T1 cells was screened by CCK-8 assay.The invasion and migration abilities of 4T1 cells following esculetin treatment at the selected drug concentra-tion were analyzed by scratch test and Transwell assay.Mitochondrial morphological change were examined by electron mi-croscopy.The levels of ferroptotic cell death were then quantified by Fe2+,reactive oxygen species(ROS),malondialde-hyde(MDA),glutathione(GSH),and GPX4 assays.Western blot was performed to evaluate the protein levels of p53,SLC7A11,GPX4 and acyl-CoA synthetase long-chain family member 4(ACSL4).RESULTS:Compared with CON group,the esculetin 1/2 IC50,IC50 and 2 IC50 groups showed smaller size,increased membrane density,and decreased ridge of mitochodria,as well as decreased levels of GSH and GPX4,reduced cell invasion and migration abilities,and de-creased levels of SLC7A11 and GPX4 proteins(P<0.05).Furthermore,these groups exhibited increased levels of Fe2+,ROS,and MDA,as well as elevated p53 and ACSL4 protein abundance(P<0.05).CONCLUSION:Esculetin pro-motes ferroptosis of 4T1 cells through a mechanism involving the p53/SLC7A11/GPX4 regulatory axis.

4.
Article in Chinese | WPRIM | ID: wpr-996499

ABSTRACT

Colorectal cancer is one of the common malignant tumors with high morbidity, and changes in lifestyle, dietary structure and environment in China in recent decades have been associated with an increase in the incidence of colorectal cancer. A large number of studies have shown that traditional Chinese medicine(TCM) can be used as a complementary and alternative treatment for colorectal cancer after conventional western medicine treatment. TCM physicians have accumulated a lot of clinical experience in the treatment of patients with stage Ⅰ-Ⅲ colorectal cancer, and have proved that TCM has unique efficacy, but there is still a lack of relevant clinical practice guidelines to standardize and guide the diagnosis and treatment of TCM. Based on this, according to the guideline development process of the World Health Organization Handbook for Guideline Development and the Clinical Evidence Grading Criteria on TCM Based on Evidence Body, under the framework of relevant laws, regulations and technical guidance documents, combined with the evidence of relevant domestic and foreign clinical research in recent years for evidence grading and opinion recommendation, and then the Guidelines for TCM Intervention After Conventional Western Medicine Treatment for Stage Ⅰ-Ⅲ Colorectal Cancer were developed by expert consensus. This guideline introduces the etiology, pathogenesis, syndrome differentiation and treatment of TCM intervention for colorectal cancer, which can provide guiding opinions for TCM clinicians and clinicians of integrated traditional Chinese and western medicine engaged in the prevention and treatment of colorectal cancer.

5.
Article in Chinese | WPRIM | ID: wpr-1014959

ABSTRACT

Liver cancer is one of the most dangerous malignant tumors in China. It has the characteristics of high mortality and occult onset. At present, the treatment of liver cancer is mainly targeted drugs, surgery and chemoradiotherapy, which is prone to high recurrence rate and strong drug resistance. In this paper, glutamine metabolism as the breakthrough point, the glutamine metabolism enzymes and related genes related to liver cancer are reviewed, in order to provide an open new idea for the pathogenesis and drug treatment of liver cancer.

6.
Article in Chinese | WPRIM | ID: wpr-462540

ABSTRACT

Objective To observe the influencing of Yiqi Huayu Jiedu Prescription on the growth of HepG2 nude mice transplantation tumor and the expression of related factors of vascular mimicry. Methods Models of transplanted tumors, which were made by HepG2 cells in nude mice, were randomly divided into 7 groups, Yiqi Huayu Jiedu Prescription group, Astragali Radix group, Curcumae Rhizoma group, Paridis Rhizoma group, Gecko group, cis-platinum group, and model group. Except for the model group, the rest groups were given relevant medicine for intervention. 21 days later. HIF-1α, MMP-2, MMP-9, and E-cad were detected by immunohistochemistry, and Twist1 and Bcl-2 were detected by fluorescence quantitative PCR. Results Compared with the model group, tumor volume in the rest groups decreased (P<0.05), and the effect in the Yiqi Huayu Jiedu Prescription group was more obvious than the Astragali Radix group, Paridis Rhizoma group and Gecko group (P<0.05);The expression of vasculogenic mimicry structure was rare in each group, and the model group and cis-platinum group were the most obvious;Except for the Astragali Radix group, the expressions of HIF-1α, MMP-2, and MMP-9 showed statistical significance compared with model group (P<0.05);The expression of E-cad in the Yiqi Huayu Jiedu Prescription group and Astragali Radix group showed statistical significance (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group, Paridis Rhizoma group, and Gecko group decreased significantly compared with the model group (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group was much better than the other groups (P<0.05);The expression of Twist1 showed statistical significance in the Yiqi Huayu Jiedu Prescription group, Curcumae Rhizoma group, Paridis Rhizoma group, and cis-platinum group (P<0.05). Conclusion Yiqi Huayu Jiedu Prescription can reduce expression of HIF-1α, Twist1, Bcl-2, MMP-2, and MMP-9, and increase expression of E-cad, thereby inhibiting the formation of vascular mimicry.

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