1.Value of Ultrasonographic Features Combined With Immunohistochemistry in Predicting Axillary Lymph Node Metastasis in Middle-Aged Women With Breast Cancer.
Qian-Kun CHANG ; Wen-Ying WU ; Chun-Qiang BAI ; Zhi-Chao DING ; Wei-Fang WANG ; Ming-Han LIU
Acta Academiae Medicinae Sinicae 2025;47(4):550-556
Objective To investigate the value of ultrasonographic features combined with immunohistochemistry in predicting axillary lymph node metastasis in middle-aged women with breast cancer.Methods A retrospective analysis was conducted on 827 middle-aged female breast cancer patients who underwent surgical treatment at the Affiliated Hospital of Chengde Medical University from June 2017 to June 2023.Ultrasonographic and immunohistochemical information was collected,and the patients were randomly allocated into a training set(579 patients)and a validation set(248 patients).Univariate and multivariate Logistic regression analyses were performed to identify ultrasonographic and immunohistochemical risk factors associated with axillary lymph node metastasis in these patients,and a nomogram model was developed.Receiver operating characteristic curves and calibration curves were established to evaluate the performance of the nomogram model,and clinical decision curves were built to assess the clinical value of the model.Results The maximum diameter,morphology,boundary,calcification,and expression of human epidermal growth facor receptor 2 and Ki-67 in breast cancer lesions were identified as risk factors for predicting axillary lymph node metastasis in middle-aged women.The areas under the curve of the nomogram model on the training and validation sets were 0.747(0.707-0.787)and 0.714(0.647-0.780),respectively.Calibration curves and clinical decision curves indicated good consistency and performance of the model.Conclusion The nomogram model constructed based on ultrasonographic features and immunohistochemistry of the primary breast cancer lesion demonstrates high value in predicting axillary lymph node metastasis in middle-aged women with breast cancer.
Humans
;
Female
;
Breast Neoplasms/diagnostic imaging*
;
Middle Aged
;
Lymphatic Metastasis/diagnostic imaging*
;
Axilla
;
Retrospective Studies
;
Nomograms
;
Ultrasonography
;
Immunohistochemistry
;
Lymph Nodes/diagnostic imaging*
;
Risk Factors
;
Ki-67 Antigen
2.Clinical significance of tertiary lymphoid structure maturity in colorectal cancer patients.
Jiangjiang ZHENG ; Jingjing YU ; Jingjing XIE ; Dong CHEN ; Hong DENG
Journal of Zhejiang University. Medical sciences 2024;53(6):765-771
OBJECTIVES:
To explore the clinical significance of the tertiary lymphoid structure (TLS) maturity in colorectal cancer patients.
METHODS:
A total of 230 surgically removed colorectal cancer specimens with detailed follow-up data were collected from Yinzhou Second Hospital. The patients were divided into mature TLS group and immature TLS group according to immunohistochemical results. The patient age, gender, maximum tumor diameter, tumor location, differentiation degree, depth of invasion, lymph node metastasis, vascular tumor thrombus, liver metastasis, distant non-liver metastasis, mismatch repair status, expression of Ki-67, P53 and programmed death-ligand (PD-L) 1 were analyzed. The Kaplan-Meier method (Breslow test) was used to analyze the survival of patients, and multivariate Cox regression model was applied to analyze the prognostic factors.
RESULTS:
There were 128 cases of mature TLS and 102 cases of immature TLS. Compared to the immature TLS group, the mature TLS group showed a significantly lower rate of vascular tumor thrombus, lymph node metastasis, and liver metastasis. Additionally, the positive expression rate of Ki-67 was markedly reduced, while the rate of deficient mismatch repair and the positive rate of PD-L1 were significantly increased (all P<0.05). The overall survival rate of the mature TLS group was superior to that of the immature TLS group (Breslow=4.553, P<0.05). Cox regression analysis indicated that lymph node metastasis was an independent risk factor for the prognosis of colorectal cancer patients (P<0.01), while TLS maturation was a protective factor (P<0.05).
CONCLUSIONS
The formation of TLS may play a significant role in inhibiting lymph node metastasis, liver metastasis, and vascular tumor thrombus in colorectal cancer. In addition, patients with mature TLS have a favorable clinical prognosis.
Humans
;
Colorectal Neoplasms/pathology*
;
Male
;
Female
;
Middle Aged
;
Tertiary Lymphoid Structures/pathology*
;
Prognosis
;
Aged
;
Adult
;
Lymphatic Metastasis
;
Ki-67 Antigen/metabolism*
;
B7-H1 Antigen/metabolism*
;
Clinical Relevance
3.Preoperative CT radiomics-based model for predicting Ki-67 expression in clear cell renal cell carcinoma patients.
Zhijun YANG ; Han HE ; Yunfeng ZHANG ; Jia WANG ; Wenbo ZHANG ; Fenghai ZHOU
Journal of Central South University(Medical Sciences) 2024;49(11):1722-1731
OBJECTIVES:
Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma (RCC), and developing personalized treatment strategies is crucial for improving patient prognosis. This study aims to develop and validate a preoperative computer tomography (CT) radiomics-based predictive model to estimate Ki-67 expression in ccRCC patients, thereby assisting in clinical treatment decisions and prognosis prediction.
METHODS:
A retrospective analysis was conducted on 214 ccRCC patients who underwent surgical treatment at Gansu Provincial Hospital between January 2018 and November 2023. Patients were classified into high Ki-67 expression (n=123) and low Ki-67 expression (n=91) groups based on postoperative immunohistochemical staining results. The dataset was randomly divided in a 7꞉3 ratio into a training set (n=149) and a validation set (n=65). Preoperative contrast-enhanced urinary CT images and clinical data were collected. After preprocessing, 5 mm arterial-phase CT images were manually segmented layer by layer to delineate the region of interest (ROI) using ITK-SNAP 3.8 software. Radiomic features were then extracted using the FeAture Explorer (FAE) package. Dimensionality reduction and feature selection were performed using the least absolute shrinkage and selection operator (LASSO) algorithm, yielding the optimal feature set. Three classification models were constructed using logistic regression (LR), multilayer perceptron (MLP), and support vector machine (SVM). The receiver operating characteristic (ROC) curve, area under the curve (AUC), decision curve analysis (DCA), and calibration curves were used for model evaluation.
RESULTS:
A total of 107 radiomic features were extracted from 5 mm arterial-phase CT images, and twenty-one features significantly associated with Ki-67 expression were selected using the LASSO algorithm. Predictive models were developed using LR, MLP, and SVM classifiers. In the training and validation sets, the AUC values for each model were 0.904 (95% CI 0.852 to 0.956) and 0.818 (95% CI 0.710 to 0.926) for the LR model, 0.859 (95% CI 0.794 to 0.923) and 0.823 (95% CI 0.716 to 0.929) for the MLP model, and 0.917 (95% CI 0.865 to 0.969) and 0.857 (95% CI 0.760 to 0.953) for the SVM model. DCA demonstrated that all models had good clinical net benefit, while calibration curves indicated high accuracy of the predictions, supporting the robustness and reliability of the models.
CONCLUSIONS
A CT radiomics-based model for predicting Ki-67 expression in ccRCC was successfully developed. This model provides valuable guidance for treatment planning and prognostic assessment in ccRCC patients.
Humans
;
Carcinoma, Renal Cell/surgery*
;
Kidney Neoplasms/surgery*
;
Tomography, X-Ray Computed/methods*
;
Ki-67 Antigen/metabolism*
;
Retrospective Studies
;
Female
;
Male
;
Middle Aged
;
Aged
;
Prognosis
;
Adult
;
Preoperative Period
;
Radiomics
4.Correlation between SWE parameters and histopathological features and immunohistochemical biomarkers in invasive breast cancer.
Xu LIU ; Jigang LI ; Ying HE ; Zhiyuan WANG
Journal of Central South University(Medical Sciences) 2024;49(12):1941-1952
OBJECTIVES:
Shear wave elastography (SWE) is a novel quantitative elastography technique that can assess the hardness of different tissues. This study introduces a novel shear wave parameter-frequency of mass characteristic (fmass)-and investigates its correlation, along with other shear wave parameters, with the histopathological features and immunohistochemical (IHC) biomarkers of invasive breast cancer (IBC). The study aims to explore whether SWE can provide useful information for IBC treatment and prognosis.
METHODS:
With the pathological results as the gold standard, 258 malignant breast lesions were collected, and all patients underwent conventional ultrasound and SWE examinations. The SWE parameters [maximum elastic value (Emax), minimum elastic value (Emin), mean elastic value (Emean), standard deviation of elastic value of the whole lesion (Esd)] and fmass] in the transverse and longitudinal orthogonal sections were measured, and their correlations with the prognostic factors of IBC [including tumor diameters, axillary lymph node (ALN) metastasis, lymphatic vessel invasion (LVI), calcification, histological type, histological grade, and IHC biomarkers (ER, PR, HER-2, Ki-67), and molecular subtypes] were analyzed. The correlations between the SWE parameters of the transverse and longitudinal sections of the tumors with different prognostic factors and the above indicators were analyzed. At the same time, the receiver operating characteristic (ROC) curve was used to analyze the efficacy of fmass in predicting ER and PR expression.
RESULTS:
Emean, Emax, Esd, and fmass were correlated with tumor diameters; Emean, Emax and Esd were correlated with histological types and histological grades. Emax and Esd were correlated with ALN metastasis, LVI and pathological types. In the IHC biomarker-labeled masses, fmass was correlated with ER and PR (both P<0.05), and Emean, Emax, and Esd were correlated with HER-2 and Ki-67 (all P<0.05). Emean, Emax, and fmass were all correlated with breast cancer subtypes (all P<0.05), and Emean and Emax were higher in Luminal B [HER-2(+)] breast cancer, while fmass was lower in HER-2(+) and triple-negative breast cancer. Among the statistically significant prognostic factors, the P values of the transverse sections of the masses were all less than or equal to those of the longitudinal sections. The AUC of fmass in the transverse sections of the masses for predicting ER and PR expression were 0.73 (95% CI 0.65 to 0.80) and 0.67 (95% CI 0.60 to 0.74), respectively, with the optimal cut-off values being 76.50 and 60.66, the sensitivities being 72.45% and 81.98%, the specificities being 66.13% and 45.35%, and the accuracies being 70.93% and 69.77%, respectively. The AUC of fmass in the longitudinal sections of the masses for predicting ER and PR expression were 0.74 (95% CI 0.67 to 0.81) and 0.65 (95% CI 0.58 to 0.72), respectively, with the optimal cut-off values being 131.8 and 137.5, the sensitivities being 69.90% and 66.28%, the specificities being 72.58% and 60.47%, and the accuracies being 70.54% and 64.34%, respectively. The fmass in the transverse sections of the masses was more statistically significant.
CONCLUSIONS
The poor prognosis factors of IBC are related to high Emean, Emin, Emax, Esd, and low fmass. The fmass can predict the expression of ER and PR, and the transverse cut data are more meaningful. SWE is helpful for predicting the invasiveness of IBC.
Humans
;
Breast Neoplasms/metabolism*
;
Female
;
Elasticity Imaging Techniques/methods*
;
Biomarkers, Tumor/metabolism*
;
Middle Aged
;
Adult
;
Prognosis
;
Immunohistochemistry
;
Neoplasm Invasiveness
;
Receptor, ErbB-2/metabolism*
;
Aged
;
Lymphatic Metastasis
;
Receptors, Estrogen/metabolism*
;
Receptors, Progesterone/metabolism*
;
Ki-67 Antigen/metabolism*
5.A multicenter study of the clinicopathological characteristics and a risk prediction model of early-stage breast cancer with hormone receptor-positive/human epidermal growth factor receptor 2-low expression.
Ling XIN ; Qian WU ; Chongming ZHAN ; Hongyan QIN ; Hongyu XIANG ; Min GAO ; Xuening DUAN ; Yinhua LIU ; Jingming YE
Chinese Medical Journal 2023;136(24):2967-2973
BACKGROUND:
In light of the significant clinical benefits of antibody-drug conjugates in clinical trials, the human epidermal growth factor receptor 2 (HER2)-low category in breast cancers has gained increasing attention. Therefore, we studied the clinicopathological characteristics of Chinese patients with hormone receptor (HR)-positive/HER2-low early-stage breast cancer and developed a recurrence risk prediction model.
METHODS:
Female patients with HR-positive/HER2-low early-stage breast cancer treated in 29 hospitals of the Chinese Society of Breast Surgery (CSBrS) from Jan 2015 to Dec 2016 were enrolled. Their clinicopathological data and prognostic information were collected, and machine learning methods were used to analyze the prognostic factors.
RESULTS:
In total, 25,096 patients were diagnosed with breast cancer in 29 hospitals of CSBrS from Jan 2015 to Dec 2016, and clinicopathological data for 6486 patients with HER2-low early-stage breast cancer were collected. Among them, 5629 patients (86.79%) were HR-positive. The median follow-up time was 57 months (4, 76 months); the 5-year disease-free survival (DFS) rate was 92.7%, and the 5-year overall survival (OS) rate was 97.7%. In total, 412 cases (7.31%) of metastasis were observed, and 124 (2.20%) patients died. Multivariate Cox regression analysis revealed that T stage, N stage, lymphovascular thrombosis, Ki-67 index, and prognostic stage were associated with recurrence and metastasis ( P <0.05). A recurrence risk prediction model was established using the random forest method and exhibited a sensitivity of 81.1%, specificity of 71.7%, positive predictive value of 74.1%, and negative predictive value of 79.2%.
CONCLUSION:
Most of patients with HER2-low early-stage breast cancer were HR-positive, and patients had favorable outcome; tumor N stage, lymphovascular thrombosis, Ki-67 index, and tumor prognostic stage were prognostic factors. The HR-positive/HER2-low early-stage breast cancer recurrence prediction model established based on the random forest method has a good reference value for predicting 5-year recurrence events.
REGISTRITATION
ChiCTR.org.cn, ChiCTR2100046766.
Humans
;
Female
;
Breast Neoplasms/diagnosis*
;
Ki-67 Antigen
;
Receptor, ErbB-2
;
Prognosis
;
Thrombosis
;
Receptors, Progesterone
6.Panax notoginseng saponins improve monocrotaline-induced pulmonary arterial hypertension in rats by inhibiting ADAM10/Notch3 signaling pathway.
Sai ZHANG ; Yun-Na TIAN ; Zheng-Yang SONG ; Xiao-Ting WANG ; Xin-Yu WANG ; Jun-Peng XU ; Lin-Bo YUAN ; Wan-Tie WANG
Acta Physiologica Sinica 2023;75(4):503-511
In this study, we investigated the effects of Panax notoginseng saponins (PNS) on pulmonary vascular remodeling and ADAM10/Notch3 pathway in pulmonary arterial hypertension (PAH). PAH rat model was established, and male Sprague Dawley (SD) rats were randomly divided into control group, monocrotaline (MCT) group and MCT+PNS group, with 10 rats in each group. Rats in the control group were intraperitoneally injected with equal volume of normal saline. Rats in the MCT group was injected intraperitoneally with 60 mg/kg MCT on the first day, and then with the same volume of normal saline every day. Rats in the MCT+PNS group was injected intraperitoneally with 60 mg/kg MCT on the first day, and then with 50 mg/kg PNS every day. The modeling time of each group lasted for 21 days. After the model was established, the mean pulmonary artery pressure (mPAP) was measured by right heart catheterization technique, the right ventricular hypertrophy index (RVHI) was calculated, the microscopic morphology and changes of pulmonary vascular wall were observed by HE and Masson staining, and the expressions of ADAM10, Notch3, Hes-1, P27, PCNA, Caspase-3 proteins and mRNA in pulmonary vascular tissue of rats were detected by Western blot and qPCR. The expression and localization of Notch3 and α-SMA were detected by immunofluorescence staining. The protein expression of ADAM10 was detected by immunohistochemical staining. The results showed that compared with the control group, mPAP, RVHI, pulmonary vessels and collagen fibers in the MCT group were significantly increased, the expressions of ADAM10, Notch3, Hes-1, and PCNA protein and mRNA were significantly increased, while the expressions of P27 and Caspase-3 protein and mRNA were decreased significantly. Compared with the MCT group, mPAP and RVHI were significantly decreased, pulmonary vessels were significantly improved and collagen fibers were significantly reduced, the expressions of protein and mRNA of ADAM10, Notch3, Hes-1, and PCNA were decreased in MCT+PNS group, but the expressions of protein and mRNA of P27 and Caspase-3 were increased slightly. The results of immunofluorescence showed that Notch3 and α-SMA staining could overlap, which proved that Notch3 was expressed in smooth muscle cells. The expression of Notch3 in the MCT group was increased significantly compared with that in the control group, while PNS intervention decreased the expression of Notch3. Immunohistochemical staining showed that compared with the control group, the amount of ADAM10 in the MCT group was increased significantly, and the expression of ADAM10 in the MCT+PNS group was decreased compared with the MCT group. These results indicate that PNS can improve the PAH induced by MCT in rats by inhibiting ADAM10/Notch3 signaling pathway.
Animals
;
Male
;
Rats
;
Caspase 3/metabolism*
;
Collagen
;
Disease Models, Animal
;
Hypertension, Pulmonary/drug therapy*
;
Monocrotaline/adverse effects*
;
Panax notoginseng/chemistry*
;
Proliferating Cell Nuclear Antigen/pharmacology*
;
Pulmonary Arterial Hypertension
;
Pulmonary Artery/metabolism*
;
Rats, Sprague-Dawley
;
Receptor, Notch3/genetics*
;
RNA, Messenger
;
Saline Solution
;
Signal Transduction
;
Saponins/pharmacology*
7.Clinicopathological and immunohistochemical analysis of maxillofacial granular cell tumor.
Zeliang SHEN ; Lihong YAO ; Hongjie JIANG ; Mao LI ; Yaling TANG
West China Journal of Stomatology 2023;41(4):414-420
OBJECTIVES:
To analyze the clinicopathological features of maxillofacial granular cell tumors (GCT) with the aid of immunohistochemical staining.
METHODS:
Seven cases of maxillofacial GCT were retrospectively collated, and the microscopic morphology of maxillofacial GCT was analyzed. The expression of S-100, neuron-specific enolase (NSE), SOX-10, CD68, actin, desmin, and Ki-67 in GCT was detected by immunohistochemical staining. The cases were observed in the follow-ups after clinical treatment.
RESULTS:
All seven GCT tumors lacked envelopes and were poorly defined. Microscopically, the sizes of the tumor cells were large and appeared with inconspicuous cell membranes, forming a syncytium-like appearance. The cytoplasm was filled with characteristic eosinophilic granules. The immunohistochemical results showed that six cases were NSE-positive, five cases were S-100-positive, seven cases were CD68-positive, five cases were SOX-10-positive, one case was actin-positive, and seven cases were desmin-negative. The Ki-67 index did not exceed 5% in all cases. In the follow-up sessions, none of the six cases presented a recurrence.
CONCLUSIONS
Maxillofacial GCT has a characteristic histological structure. Immunohistochemical S-100, CD68, and other indicators can assist in diagnosis, and the prognosis is good after clinical resection.
Humans
;
Ki-67 Antigen/metabolism*
;
Granular Cell Tumor/surgery*
;
Retrospective Studies
;
Actins/metabolism*
;
Desmin/metabolism*
;
S100 Proteins/metabolism*
8.Forkhead Box M1 Regulates the Proliferation,Invasion,and Drug Resistance of Gastric Cancer Cells via circ_NOTCH1.
Ning GE ; Yuan-Yuan JIANG ; Zhong-Ping PAN ; Jie WAN
Acta Academiae Medicinae Sinicae 2023;45(5):713-720
Objective To investigate the impacts of forkhead box M1(FOXM1)on the proliferation,invasion,and drug resistance of gastric cancer cells by regulating the circular RNA circ_NOTCH1.Methods Western blotting and real-time quantitative PCR were performed to determine the expression of FOXM1 protein and circ_NOTCH1,respectively,in the gastric cancer tissue,para-carcinoma tissue,human normal gastric mucosa epithelial cell line GES-1 and gastric cancer cell lines MGC-803,HGC-27,and BGC-823.BGC-823 cells were classified into the following groups:control,short hairpin RNA FOXM1(sh-FOXM1)and negative control(sh-NC),small interfering RNA circ_NOTCH1(si-circ_NOTCH1)and negative control(si-NC),and sh-FOXM1+circ_NOTCH1 overexpression plasmid(sh-FOXM1+pcDNA-circ_NOTCH1)and sh-FOXM1+negative control(sh-FOXM1+pcDNA).CCK-8 assay and clone formation assay were employed to measure the cell proliferation,and Transwell assay to measure cell invasion.After treatment with 1.0 mg/L adriamycin for 48 h,the cell resistance in each group was analyzed.Western blotting was employed to determine the expression levels of FOXM1,proliferating cell nuclear antigen(PCNA),Bax,multi-drug resistance-associated protein 1(MRP1),and multi-drug resistance gene 1(MDR1).RNA pull-down and RNA immunoprecipitation were employed to examine the binding of circ_NOTCH1 to FOXM1 protein.Results Compared with those in the para-carcinoma tissue,the expression levels of FOXM1 protein and circ_NOTCH1 in the gastric cancer tissue were up-regulated(all P<0.001).Compared with GES-1 cells,MGC-803,HGC-27,and BGC-823 cells showed up-regulated expression levels of FOXM1 protein and circ_NOTCH1(all P<0.001).Compared with the control group and sh-NC group,the sh-FOXM1 group with down-regulated expression of FOXM1 protein and circ_NOTCH1 showed decreased optical density value,clone formation rate,cell invasion number,and cell viability,down-regulated expression of PCNA,MRP1,and MDR1,and up-regulated expression of Bax protein in BGC-823 cells(all P<0.001).Compared with the control group and the si-NC group,the si-circ_NOTCH1 group with down-regulated expression of circ_NOTCH1 showed decreased optical density value,clone formation rate,cell invasion number,and cell viability,down-regulated expression of PCNA,MRP1,and MDR1,and up-regulated expression of Bax protein in BGC-823 cells(all P<0.001).Compared with sh-FOXM1 group and sh-FOXM1+pcDNA group,the sh-FOXM1+pcDNA-circ_NOTCH1 group with up-regulated expression of circ_NOTCH1 showed increased optical density value,clone formation rate,cell invasion number,and cell viability,up-regulated expression of PCNA,MRP1,and MDR1,and down-regulated expression of Bax protein(all P<0.001).FOXM1 protein was able to interact with circ_NOTCH1.Conclusion Interference with FOXM1 may inhibit the proliferation,invasion,and drug resistance of gastric cancer cells by silencing circ_NOTCH1 expression.
Humans
;
bcl-2-Associated X Protein/metabolism*
;
Carcinoma
;
Cell Line, Tumor
;
Cell Proliferation/genetics*
;
Drug Resistance
;
Forkhead Box Protein M1/metabolism*
;
Gene Expression Regulation, Neoplastic
;
MicroRNAs/genetics*
;
Proliferating Cell Nuclear Antigen/metabolism*
;
Receptor, Notch1/metabolism*
;
RNA, Small Interfering/genetics*
;
Stomach Neoplasms/genetics*
9.Association of Serine/Threonine Phosphoprotein Phosphatase 4C Expression With Prognosis of Gastric Cancer.
Zhi-Jun GENG ; Ju HUANG ; Qing-Qing LI ; Zhi-Xuan ZHOU ; Jing LI ; Xiao-Feng ZHANG ; Lian WANG ; Yue-Yue WANG ; Xue SONG ; Lu-Gen ZUO
Acta Academiae Medicinae Sinicae 2023;45(5):721-729
Objective To investigate the expression level of serine/threonine phosphoprotein phosphatase 4C(PPP4C)in gastric cancer,and analyze its relationship with prognosis and the underlying regulatory mechanism.Methods The clinical data of 104 gastric cancer patients admitted to the First Affiliated Hospital of Bengbu Medical College between January 2012 and August 2016 were collected.Immunohistochemical staining was employed to determine the expression levels of PPP4C and Ki-67 in the gastric cancer tissue.The gastric cancer cell lines BGC823 and HGC27 were cultured and transfected with the vector for PPP4C knockdown,the vector for PPP4C overexpression,and the lentiviral vector(control),respectively.The effects of PPP4C on the cell cycle and proliferation were analyzed and the possible regulatory mechanisms were explored.Results PPP4C was highly expressed in gastric cancer(P<0.001),and its expression promoted malignant progression of the tumor(all P<0.01).Univariate and Cox multivariate analysis clarified that high expression of PPP4C was an independent risk factor affecting the 5-year survival rate of gastric cancer patients(P=0.003).Gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis suggested that PPP4C may be involved in the cell cycle.The correlation analysis showed that the expression of PPP4C was positively correlated with that of Ki-67 in gastric cancer(P<0.001).The up-regulation of PPP4C expression increased the proportion of tumor cells in the S phase,alleviated the G2/M phase arrest,and promoted the proliferation of gastric cancer cells and the expression of cyclin D1 and cyclin-dependent kinase 6(CDK6)(all P<0.05).The down-regulation of PPP4C decreased the proportion of gastric cancer cells in the S phase,promoted G2/M phase arrest,and inhibited cell proliferation and the expression of cyclin D1,CDK6,and p53(all P<0.05).p53 inhibitors promoted the proliferation of BGC823 and HGC27 cells in the PPP4C knockdown group(P<0.001,P<0.001),while p53 activators inhibited the proliferation of BGC823 and HGC27 cells in the PPP4C overexpression group(P<0.001,P=0.002).Conclusions PPP4C is highly expressed in gastric cancer and affects the prognosis of the patients.It may increase the proportion of gastric cancer cells in the S phase and alleviate the G2/M phase arrest by inhibiting p53 signaling,thereby promoting cell proliferation.
Humans
;
Stomach Neoplasms/genetics*
;
Cyclin D1/metabolism*
;
Tumor Suppressor Protein p53
;
Phosphoproteins/metabolism*
;
Ki-67 Antigen
;
Cell Line, Tumor
;
Prognosis
;
Cell Proliferation
;
Phosphoprotein Phosphatases/metabolism*
;
Threonine
;
Serine
10.Blaps rynchopetera combined with cyclophosphamide affects proliferation and apoptosis of lung cancer cells via Wnt/β-catenin signaling pathway.
Jing-Nan YAN ; Ke MA ; Wen-Jie LIU ; Ying LIN ; Xiu-Yu LI ; Dan WU
China Journal of Chinese Materia Medica 2023;48(20):5603-5611
This study aims to investigate the effects of Blaps rynchopetera Fairmaire and/or cyclophosphamide on the proliferation and apoptosis of lung cancer cells and decipher the underlying mechanism. B. rynchopetera and cyclophosphamide-containing serum and blank serum were prepared from SD rats. Cell counting kit-8(CCK-8) assay was employed to examine the proliferation of lung cancer cell lines A549 and Lewis treated with corresponding agents. The Jin's formula method was used to evaluate the combined effect of the two drugs. According to the evaluation results, appropriate drug concentrations and lung cancer cell line were selected for subsequent experiments, which included control, B. rynchopetera, cyclophosphamide, B. rynchopetera + cyclophosphamide, and B. rynchopetera + Wnt/β-catenin pathway agonist lithium chloride(LiCl) groups. Immunocytochemistry was employed to measure the expression of proliferation-related proteins in Lewis cells after drug interventions. Flow cytometry was employed to determine the cell cycle and apoptosis. The expression levels of proliferating cell nuclear antigen(PCNA), cyclinD1, B-cell lymphoma 2(Bcl-2), Bcl-2-assiocated X protein(Bax), Wnt1, and β-catenin were determined by Western blot. The results showed that B. rynchopetera and/or cyclophosphamide significantly inhibited the proliferation of A549 and Lewis cells. Compared with B. rynchopetera alone, the combination increased the inhibition rate on cell proliferation. The combination of B. rynchopetera and cyclophosphamide demonstrated a synergistic effect according to Jin's formula-based evaluation. Compared with the control group, the B. rynchopetera, cyclophosphamide, and B. rynchopetera + cyclophosphamide groups showed increased proportion of Lewis cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the cyclophosphamide group, the combination group showed increased proportion of cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the B. rynchopetera group, the B. rynchopetera + LiCl group had deceased proportion of cells in G_0/G_1 phase, decreased apoptosis rate, down-regulated expression of Bax, and up-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. The results indicated that B. rynchopetera could inhibit the proliferation, arrest the cell cycle, and induce the apoptosis of lung cancer cells by inhibiting the Wnt/β-catenin signaling pathway. Moreover, B. rynchopetera had a synergistic effect with cyclophosphamide.
Rats
;
Animals
;
Wnt Signaling Pathway
;
Lung Neoplasms/genetics*
;
beta Catenin/metabolism*
;
Proliferating Cell Nuclear Antigen
;
bcl-2-Associated X Protein/metabolism*
;
Rats, Inbred Lew
;
Rats, Sprague-Dawley
;
Apoptosis
;
Proto-Oncogene Proteins c-bcl-2/metabolism*
;
Cell Proliferation
;
Cyclophosphamide
;
Cell Line, Tumor

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