Protective Effect of Qihong Tongluo Prescription on Vascular Endothelial Cells in Rats with Deep Venous Thrombosis Based on NF-κB Pathway / 中国实验方剂学杂志

ObjectiveTo explore the protective effect and mechanism of Qihong Tongluo prescription on vascular endothelial cells in rats with deep venous thrombosis （DVT）. MethodSixty-six SD rats were randomly divided into a blank group （n=11） and a modeling group （n=55）. The DVT model was induced in rats of the modeling group by slowing down blood flow and damaging vascular endothelium. The model rats were randomly divided into model group， aspirin group （200 mg·kg-1）， and low-，medium-， and high-dose Qihong Tongluo prescription groups （6.5， 13， 26 g·kg-1） according to a random number table. Rats were treated with corresponding drugs by gavage， while those in the model group and the blank group received normal saline， once per day for 7 days. The rats were sacrificed and the abdominal aortic blood was taken. The levels of serum endothelin-1 （ET-1） and interleukin-6 （IL-6） were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in vascular endothelial tissues. The ultrastructure of vascular endothelial cells was observed by the transmission electron microscope. The viability of vascular endothelial cells was detected by methylthiazolyldiphenyl-tetrazolium bromide （MTT） method，and the release level of lactate dehydrogenase （LDH） was detected by the LDH kit. The messenger ribonucleic acid （mRNA） expression of platelet-activating factor （PAF），nuclear transcription factor κB （NF-κB），Ras-related C3 botulinum toxin substrate 1 （Rac1）， and Ras-related C3 botulinum toxin substrate 2 （Rac2） in vascular endothelial tissues were detected by real-time reverse transcription polymerase chain reaction （Real-time PCR）. The protein expression of PAF，NF-κB，Rac1， and Rac2 in vascular endothelial tissues was detected by Western blot. ResultThe model group showed seriously damaged and swollen vascular endothelial cells with massive shedding， attachment of massive inflammatory cells， nucleus pyknosis and deformation under the electron microscope， highly swollen mitochondria， serious cytoplasmic vacuolation，and exposure of internal elastic membrane. The damage of vascular endothelium and its ultrastructure in Qihong Tongluo prescription groups and the aspirin group was improved in varying degrees. Compared with the blank group，the model group showed increased levels of serum ET-1 and IL-6，potentiated vascular endothelial cell viability， up-regulated mRNA and protein expression of PAF，NF-κB，Rac1， and Rac2 in vascular endothelial tissues，and decreased LDH release level of vascular endothelial cells （P<0.05）. Compared with the model group，the aspirin group and the Qihong Tongluo prescription groups showed decreased levels of serum ET-1 and IL-6，blunted vascular endothelial cell viability，down-regulated mRNA and protein expression of PAF，NF-κB，Rac1， and Rac2 in vascular endothelial tissues，and increased LDH release level of vascular endothelial cells （P<0.05）. The effect of Qihong Tongluo prescription was dose-dependent. ConclusionQihong Tongluo prescription has a protective effect on vascular endothelial cells of DVT rats and can prevent and treat thrombosis，and its therapeutic effect is presumably achieved by inhibiting the expression of PAF，NF-κB，Rac1，and Rac2 and reducing the levels of serum ET-1 and IL-6.

Rac1 promotes the formation of heterotypic cell-in-cell structure / 生物工程学报

Heterotypic cell-in-cell structures (heCICs) are closely related to tumor development and progression, and have become a new frontier in life science research. Ras-related C3 botulinum toxin substrate 1 (Rac1) belongs to the classic Rho GTPase, which plays a key role in regulating the cytoskeleton and cell movement. To investigate the role and mechanism of Rac1 in the formation of heCICs, tumor cells and immune killer cells were labeled with cell-tracker, respectively, to establish the heCICs model. Upon treatment with the Rac1 inhibitor NSC23766, the formation of heCICs between tumor and immune cells was significantly reduced. The plasmid pQCXIP-Rac1-EGFP constructed by gene cloning was packaged into pseudoviruses that subsequently infect tumor cells to make cell lines stably expressing Rac1. As a result, the formation of heCICs was significantly increased upon Rac1 overexpression. These results demonstrated a promotive role of Rac1 in heCICs formation, which may facilitate treating cell-in-cell related diseases, such as tumors, by targeting Rac1.

Expression of miR-4695-5p in the serum of colorectal cancer patients and the mechanism of its effect on the proliferation and invasion of colorectal cancer cells / 国际外科学杂志

Objective:To explore the expression of microRNA-4695-5p in the serum of colorectal cancer patients and its effect on the proliferation and invasion of colorectal cancer CACO-2 cells.Methods:A total of 43 serum samples of colorectal cancer patients who were admitted to the Department of Gastrointestinal Surgery, Luoyang Central Hospital Affiliated to Zhengzhou University from March 2018 to November 2021 were selected, and serum samples of 43 healthy people who underwent outpatient physical examination were used as controls. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels in the serum of colorectal cancer patients and those of healthy individuals. The miR-4695-5p overexpression plasmid or the negative control plasmid were transfected into CACO-2 cells, namely the miR-4695-5p group and the control group, and the transfection efficiency was verified by qRT-PCR. CCK8 method and Transwell experiment were used to detect the effect of overexpression of miR-4695-5p on the proliferation and invasion of CACO-2 cells. The dual luciferase reporter gene experiment was used to verify the targeted binding relationship between miR-4695-5p and Ras-related C3 botulinum toxin substrate 1 (RAC1). qRT-PCR and Western blot were used to detect the effect of overexpression of miR-4695-5p on the expression of RAC1 gene and Wnt/β-Catenin signaling pathway protein.The software of SPSS28.0 was used to conduct data analysis. The measurement data of normal distribution were espressed by Mean±SD. The t-test was used to compare the means between two groups, and the one-way analysis of variance was used to compare the means of multiple groups. Results:The expression level of miR-4695-5p in the serum of patients with colorectal cancer was significantly lower than that of healthy individuals ( P<0.01). The relative expression levels of miR-4695-5p in the control group and miR-4695-5p group were 1.09 ± 0.65 and 8.83±2.03, respectively. The expression level of miR-4695-5p in CACO-2 cells in the miR-4695-5p group was 8.10 times that of the control group, and CACO-2 cells overexpressing miR-4695-5p were successfully constructed ( P<0.01). Compared with the control group, the proliferation ability of CACO-2 cells in the miR-4695-5p group was significantly reduced ( P<0.05), and the invasion ability of CACO-2 cells was significantly reduced ( P<0.01). Bioinformatics tools and dual luciferase reporter gene experiments confirmed that miR-4695-5p can target and bind RAC1 ( P<0.01). Compared with the control group, the expression of RAC1 gene in the miR-4695-5p group was significantly decreased ( P<0.01), and the expression of Wnt/β-Catenin signaling pathway proteins Wnt3a, β-catenin, and c-MYC decreased significantly. Conclusions:miR-4695-5p is lowly expressed in the serum of colorectal cancer patients. miR-4695-5p inhibits the proliferation and invasion of colorectal cancer CACO-2 cells by targeting the inhibition of RAC1 gene expression and down-regulating the activity of the Wnt/β-Catenin signaling pathway.

Effect of PSD-95 on long-term learning and memory impairment in neonatal rats induced by sevoflurane anesthesia / 中华行为医学与脑科学杂志

Objective:To investigate the effect of postsynaptic density-95(PSD-95)on long-term learning and memory impairment in neonatal rats induced by sevoflurane anesthesia.Methods:A total of 54 SD rats aged 7 days of SPF grade were randomly divided into 3 groups: control group (exposed to air), model group (exposed to 2.1% sevoflurane, 4 h/d, consecutive 3 days) and PSD-95 inhibitor group (inhaled sevoflurane+ intraperitoneal injection NA-1, consecutive 5 days), with 18 rats in each group.Morris water maze test and new object recognition test were used to detect the ability of visuospatial learning and memory and recognition memory of rats in each group.RT-qPCR was used to detect the mRNA levels of kalirin, Rac1 and PSD-95 in rat hippocampus.The expressions of kalirin, Rac1, PSD-95 and apoptosis related proteins Caspase-3, Bcl-2 and Bax in rat hippocampus were detected by Western blot.The expression levels of kalirin and Rac1 in hippocampus were detected by immunohistochemistry.SPSS 23.0 software was used for statistical analysis.Repeated measurement ANOVA and one-way ANOVA was used for comparing among groups.Results:Repeated measurement ANOVA showed that in the water maze test, the interaction between time and group of platform seeking latency and swimming distance of the three groups were significant ( Ftime×group=36.539, 41.548, both P<0.01). Simple effect analysis showed that the platform latency and swimming distance in the model group from day 2 to 6 were longer than those in the control group (platform latency from day 2 to 6: t=14.039, 17.147, 13.155, 13.831, 27.247, all P<0.01; swimming distance from day 2 to 6: t=10.122, 20.987, 7.267, 10.011, 8.121, all P<0.01). Compared with the model group, from day 2 to 6, the platform latencies of PSD-95 inhibitor group were prolonged( t=7.948, 14.768, 11.582, 12.832, 24.346, all P<0.01) and the swimming distances were increased( t=8.235, 24.325, 11.234, 12.031, 7.036, all P<0.01). The new object recognition test found that the new object exploration time in the model group was significantly longer than that in the control group ((21.30±2.27)s, (19.21±1.42)s, t=1.843, P<0.01), and the new object exploration time in the PSD-95 inhibitor group was significantly longer than that in the model group ((26.83±2.13)s, t=4.844, P<0.01). The difference index of novel objects in the model group was significantly lower than that in the control group ((0.41±0.12), (0.59±0.10), t=3.416, P<0.01), and the difference index of novel objects in the PSD-95 inhibitor group was significantly lower than that in the model group ((0.37±0.08), t=0.696, P<0.05). The qRT-PCR results showed that the expressions of Rac1, kalirin and PSD-95 mRNA in the model group were significantly lower than those in the control group ( t=9.969, 3.954, 6.561, P<0.05), and the expressions of Rac1, kalirin and PSD-95 mRNA in the PSD-95 inhibitor group were significantly lower than those of the model group ( t=2.132, 2.251, 3.502, all P<0.05). Immunohistochemistry results showed that the kalirin in the hippocampus CA1 area of the model group was significantly lower than that in the control group((8.18±1.94) vs (15.47±3.35), t=11.47, P<0.01), and kalirin in the PSD-95 inhibitor group was significantly lower than that in the model group((4.98±1.53), t=10.28, P<0.01); Rac1 in the model group was significantly lower than that in the control group ((3.72±1.53), (8.17±2.91), t=6.76, P<0.01), and the Rac1 in the PSD-95 inhibitor group(2.73±0.37) was significantly lower than the model group ( t=4.72, P<0.05). Western blot results showed that Caspase-3((1.37±0.16) vs (0.54±0.01), t=5.71, P<0.01) and Bax((1.87±0.31) vs (1.23±0.25), t=12.01, P<0.01) protein levels in the model group were significantly higher than those in the control group.Caspase-3 and Bax protein levels in the PSD-95 inhibitor group were significantly higher than those in model group (Caspase-3: (1.79±0.17), t=9.87, P<0.01; Bax: (2.19±0.21), t=16.19, P<0.01). The Bcl-2 protein level in the model group was significantly lower than that of the control group ((1.22±0.21) vs (1.96±0.38), t=11.92, P<0.01). And the Bcl-2 protein level in the PSD-95 inhibitor group (1.01±0.19) was significantly lower than that in the model group ( t=10.73, P<0.01). Conclusion:Sevoflurane anesthesia can damage the long-term learning and memory function and reduce the expression of PSD95 protein in neonatal rats.Inhibiting the expression of PSD95 can aggravate this damage, which may be related to the synaptic plasticity and apoptosis of neurons involved in PSD95.

The Neural Mechanisms Underlying Forgetting and Its Correlations with Nervous System Diseases / 中国生物化学与分子生物学报

Forgetting is a critical component of the memory system.On the one hand, under physiological conditions, normal forgetting helps maintain the homeostasis of the brain memory system; on the other hand, abnormal forgetting is closely related to the occurrence and development of memory- related disorders under various neural pathological conditions.In another word, forgetting is for better memory.Forgetting of unpleasant or unnecessary memories is beneficial to update new information to adapt organisms to changing environment.Abnormal forgetting is usually associated with memory-related disorders.For example, patients with Alzheimer' s disease (AD) and epilepsy have symptoms of accelerated forgetting, and post-traumatic stress disorder (PTSD) and autistic patients cannot forget unpleasant memories.Currently, the essential relation and distinction between normal forgetting under physiological conditions and abnormal forgetting under pathological conditions are still unclear, and how to improve the memory impairment of patients by regulating the forgetting proeess remains to be further studied.This review mainly foeuses on the involvement of Ras-related C3 botulinum toxin substrate 1 (Racl) , eell division cyele 42 (Cde42) , neurogenesis, microglia, dopamine and a-amino-3-hydroxy-5- methyl-4-isoxazolepropionic aeid receptors (AM PA receptors) in the regulation of forgetting under physiological conditions, and abnormal forgetting in various central pathological conditions such as AD, epilepsy, PTSD and autism, which will provide insight to the neuromolecular mechanism of forgetting and new ideas for the prevention and treatment of memory-related diseases.

miR-223-3p regulates proliferation and apoptosis of hepatocellular carcinoma SMMC-7721 cells by targeting RAC1 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of miR-223-3p on the proliferation and apoptosis of hepatocellular carcinoma (HCC) cells by regulating Ras-related C3 botulinum toxin substrate 1 (RAC1) and its possible mechanism. Methods: Thirty pairs of HCC and corresponding para-cancer tissues resected in Jilin Central Hospital from August 2016 to August 2018 were collected for this study; in addition, human HCC cell lines SMMC-7721, BEL-7402, HepG2 and human normal hepatocyte QSG-7701 were also collected. The expression level of miR-223-3p in HCC tissue and cell lines was detected by qPCR. miR-223-3p mimics, miR-223-3p inhibitor and siRAC1 were transfected into SMMC-7221 cells, respectively. CCK-8 assay, Colony formation assay and Annexin V-FITC/PI staining Flow cytometry were used to detect the proliferation, clone formation and apoptosis of SMMC-7721 cells, respectively. The relationship between miR-223-3p and RAC1 was confirmed by Dual luciferase reporter gene assay. The protein level of RAC1 in SMMC-7721 cells was detected by Western blotting. Results: The expression of miR-223-3p in HCC tissues was significantly lower than that in paracaner tissues (P<0.01), and had significant correlation with pathological characteristics, such as tumor size, TNM stage, EdmondsonSteiner grade (all P<0.05 or P<0.01). miR-223-3p expression in HCC cell lines was significantly lower than that in QSG-7701 cells with the lowest expression in SMMC-7721 cells. Dual luciferase reporter gene assay confirmed that RAC1 was a target gene of miR-223-3p, and miR-223-3p negatively regulated RAC1 expression. Over-expression of miR-223-3p significantly inhibited the proliferation and colony formation (P<0.05 or P<0.01) of SMMC-7721 cells and promoted cell apoptosis (P<0.01). Contrarily, knockdown of miR-223-3p reversed the inhibitory effect of miR-223-3p mimics on cells. Conclusion: miR-223-3p over-expression inhibits proliferation and colony formation and promotes apoptosis of HCC cells, the mechanism of which may be related with its targeted down-regulation of RAC1.

Inhibition of chemotherapy-related breast tumor EMT by application of redox-sensitive siRNA delivery system CSO-ss-SA/siRNA along with doxorubicin treatment / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Metastasis is one of the main reasons causing death in cancer patients. It was reported that chemotherapy might induce metastasis. In order to uncover the mechanism of chemotherapy-induced metastasis and find solutions to inhibit treatment-induced metastasis, the relationship between epithelial-mesenchymal transition (EMT) and doxorubicin (DOX) treatment was investigated and a redox-sensitive small interfering RNA (siRNA) delivery system was designed. DOX-related reactive oxygen species (ROS) were found to be responsible for the invasiveness of tumor cells in vitro, causing enhanced EMT and cytoskeleton reconstruction regulated by Ras-related C3 botulinum toxin substrate 1 (RAC1). In order to decrease RAC1, a redox-sensitive glycolipid drug delivery system (chitosan-ss-stearylamine conjugate (CSO-ss-SA)) was designed to carry siRNA, forming a gene delivery system (CSO-ss-SA/siRNA) downregulating RAC1. CSO-ss-SA/siRNA exhibited an enhanced redox sensitivity compared to nonresponsive complexes in 10 mmol/L glutathione (GSH) and showed a significant safety. CSO-ss-SA/siRNA could effectively transmit siRNA into tumor cells, reducing the expression of RAC1 protein by 38.2% and decreasing the number of tumor-induced invasion cells by 42.5%. When combined with DOX, CSO-ss-SA/siRNA remarkably inhibited the chemotherapy-induced EMT in vivo and enhanced therapeutic efficiency. The present study indicates that RAC1 protein is a key regulator of chemotherapy-induced EMT and CSO-ss-SA/siRNA silencing RAC1 could efficiently decrease the tumor metastasis risk after chemotherapy.

Inhibition of chemotherapy-related breast tumor EMT by application of redox-sensitive siRNA delivery system CSO-ss-SA/siRNA along with doxorubicin treatment / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Metastasis is one of the main reasons causing death in cancer patients. It was reported that chemotherapy might induce metastasis. In order to uncover the mechanism of chemotherapy-induced metastasis and find solutions to inhibit treatment-induced metastasis, the relationship between epithelial-mesenchymal transition (EMT) and doxorubicin (DOX) treatment was investigated and a redox-sensitive small interfering RNA (siRNA) delivery system was designed. DOX-related reactive oxygen species (ROS) were found to be responsible for the invasiveness of tumor cells in vitro, causing enhanced EMT and cytoskeleton reconstruction regulated by Ras-related C3 botulinum toxin substrate 1 (RAC1). In order to decrease RAC1, a redox-sensitive glycolipid drug delivery system (chitosan-ss-stearylamine conjugate (CSO-ss-SA)) was designed to carry siRNA, forming a gene delivery system (CSO-ss-SA/siRNA) down-regulating RAC1. CSO-ss-SA/siRNA exhibited an enhanced redox sensitivity compared to nonresponsive complexes in 10 mmol/L glutathione (GSH) and showed a significant safety. CSO-ss-SA/siRNA could effectively transmit siRNA into tumor cells, reducing the expression of RAC1 protein by 38.2% and decreasing the number of tumor-induced invasion cells by 42.5%. When combined with DOX, CSO-ss-SA/siRNA remarkably inhibited the chemotherapy-induced EMT in vivo and enhanced therapeutic efficiency. The present study indicates that RAC1 protein is a key regulator of chemotherapy-induced EMT and CSO-ss-SA/siRNA silencing RAC1 could efficiently decrease the tumor metastasis risk after chemotherapy.

Study on Rac1/MAPK/ERK pathway mediated mechanism and role in rats with ventilator induced lung injury / 中华危重病急救医学

Objective To investigate the role of Ras-related C3 botulinum toxin substrate 1/mitogen-activated protein kinase/extracellular signal-regulated kinase (Rac 1/MAPK/ERK) signal pathway in rats with ventilator induced lung injury (VILI) and its mechanism.Methods Thirty Sprague-Dawley (SD) rats were randomly divided into spontaneous respiration group,normal tidal volume (VT) group and high VT group with 10 rats in each group.The rats in spontaneous respiration group were kept their spontaneous breathing.The rats in normal VT group and high VT group were performed tracheal intubation after tracheostomy,and underwent mechanical ventilation on bilateral lungs with 6 mL/kg and 40 mL/kg VT respectively with maintenance anesthesia.After 4-hour ventilation,heart blood,bronchoalveolar lavage fluid (BALF) and lung tissues were harvested.The levels of interleukins (IL-1β,IL-6),tumor necrosis factor-α (TNF-α),myeloperoxidase (MPO) and macrophage inflammatory protein-2 (MIP-2) in serum and BALF were determined by enzyme linked immunosorbent assay (ELISA).Lung wet/dry radio (W/D) was determined.The lung tissues were stained with hematoxylin and eosin (HE),and pathological changes were observed,and pathological scores were evaluated.The ultra structure changes in type Ⅱ alveolar epithelial cells (AEC Ⅱ)were observed with transmission electron microscope.The positive expressions of phosphorylation of extracellular signal-regulated kinase (p-ERK) were determined by immunohistochemistry,and those of Racl and F-actin were determined by immunofluorescence.The mRNA expressions of ERK and Rac1 were determined by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR),and protein expressions of Rac-1,p-ERK and F-actin were determined by Western Blot.Results ① Compared with spontaneous breathing group,lung W/D in both mechanical ventilation groups was significantly increased,with more significant increase in the high VT group (6.64 ± 0.88 vs.1.79 ± 0.36,P ＜ 0.01).② There was no obvious pathological changes in the lung tissue and AEC Ⅱ of the spontaneously breathing group.In the normal VT group,there was slight edema and infiltration of inflammatory cells;AEC Ⅱ had less lamellar bodies and uniform distribution of the villi of the alveolar epithelium.In the high VT group,the edema of the lung tissue,the widening of the pulmonary septum,the alveolus congestion,the infiltration of inflammatory cells,and alveolar structure disorder were found;and AEC Ⅱ was irregular,the number of lamellar bodies in the plastids was decreased and was unevenly distributed.The pulmonary histopathological score in the high VT group was significantly higher than that in the spontaneous breathing group and the normal VT group (12.00 ± 2.00 vs.6.00 ± 1.51,8.50 ± 0.53,both P ＜ 0.01).③ Compared with spontaneous breathing group,IL-1β,IL-6,TNF-α,MPO,and MIP-2in serum and BALF in both mechanical ventilation groups were significantly increased,with more siguificant increase in the high VT group [serum IL-1 β (ng/L):104.2 ± 15.1 vs.20.3 ± 8.3,IL-6 (ng/L):46.6 ± 11.5 vs.22.7 ± 7.5,TNF-α (ng/L):39.4±6.5 vs.5.4± 1.9,MPO (ng/L):0.66±0.24 vs.0.06±0.03,MIP-2 (ng/L):109.2±25.8 vs.22.8±8.4;BALF IL-1 β (ng/L):121.5 ± 25.6 vs.24.0 ± 7.5,IL-6 (ng/L):136.7 ± 32.7 vs.31.4 ± 10.5,TNF-α (ng/L):98.0 ± 14.8vs.10.1 ±2.6,MPO (ng/L):0.80±0.31 vs.0.08±0.04,MIP-2 (ng/L):144.4±28.9 vs.41.2±20.7;all P ＜ 0.01].④ There were only a few p-ERK,Rac1 and F-actin positive expressions in the spontaneous breathing group.The positive expressions in normal VT group were increased.In high VT group,the positive expression of p-ERK was significantly increased;Rac1 and F-actin were mainly distributed in the cell membrane and cytoplasm respectively,the positive expressions were further enhanced.⑤ The gene expressions of ERK and Rac1,and protein expressions of p-ERK,Rac1 and F-actin in the high VT group were significantly higher than those in the spontaneous breathing group and normal VT group [ERK mRNA (2-△△Ct):8.23±2.83 vs.1,3.02± 1.38,p-ERK protein (gray value):1.15±0.36 vs.0.61 ±0.23,0.88±0.22;Rac1 mRNA (2-△△Ct):4.45 ±2.26 vs.1,1.22±0.39,Rac1 protein (gray value):0.91 ±0.16 vs.0.48±0.11,0.55 ± 0.10;F-actin protein (gray value):0.70± 0.09 vs.0.49 ± 0.08,0.55 ± 0.04;all P ＜ 0.01].Conclusion F-actin expression in lung tissue was up-regulated in rats with VILI,which resulted in reconstruction of AEC Ⅱ cyto-skeleton,and variation of cell membrane permeability through Rac 1 /MAPK/ERK sigualing pathway during VILI.

Expression and clinical significance of Rac-1 and HIF-1α in breast cancer / 肿瘤研究与临床

ObjectiveTo explore the expression and clinical significance of Ras-related C3 botulinum toxin substrate 1 (Rac-1) protein and hypoxia-inducible factor 1α (HIF-1α) in breast cancer and their relationship with tumor stage, node metastasis status and hormone receptor status. MethodsImmunohistochemical method was used to detect the expression of Rac-1 and HIF-1α protein in 139 specimens of breast cancer,combined with their clinical pathological characteristics and hormone receptor status for statistical analysis.Results The expression of Rac-1 and HIF-1α in breast cancer tissue were 64.75 ％ and 65.47 ％,respectively.The levels were related with the stage of TNM and histological grade (P ＜ 0.05,P ＜ 0.01).There was no significant correlation between these two proteins'expression and the age of patient,primary pathological location, histologic type and menstruation status of breast cancer patients(P ＞ 0.05). The expression of Rac-1 and HIF-1α had no statistical significance between triple negative breast cancer(TNBC)and non-triple negative breast cancer (NTNBC) patients (x2 =1.1229,x2 =0.1786,P ＞ 0.05); The expression level of Rac-1 was positively correlated with HIF-1α in these breast cancer specimens (rs =0.414,P ＜ 0.01).ConclusionThe expression of Rac-1 is positively correlated with HIF-1α level in breast cancer.The relationship between Rac-1 and HIF-1α might be as a new important marker to estimate the invasion,metastasis extent and prognosis in breast cancer.Blocking the regulation between Rac-1 and HIF-1α might be a new treatment target in breast cancer.

The relationship between Rac1 mRNA gene levels detected by FQ-RT-PCR and the metastasis in gastric carcinomas / 中华检验医学杂志

Objective To investigate the relationship between tumor metastasis-related Rac1 mRNA expression levels and gastric carcinomas metastasis, and to investigate the significance of Rac1 as a tumor marker for the evaluation of gastric carcinomas metastasis and distinguish between benign and malignant lesions. Methods This experiment used fluorescence quantitative RT-PCR TaqMan probe technology, chose Rac1 target gene fragment, which was cloned into the pET-20b (+) vector, constructed recombinant plasmid, and established the Rac1 mRNA fluorescence quantitative RT-PCR standard curve. And then the Rac1 mRNA levels were detected in 52 cases of gastric carcinoma tissues,52 cases of para-carcinoma tissues and 12 cases of benign gastric disease tissues. Its association with the metastasis of gastric carcinomas was analyzed. Results The positive rates and levels (median, P25-P75) of Rac1 mRNA were 100. 0% ,7.41 ×105 (3. 50 × 105-4. 36 × 106) copies/μl in gastric carcinomas, 46. 2% ,0(0-1.73 × 104) copies/μl in parscarcinoma tissues, 33. 3%, 0(0-3.55 × 103) copies/μl in benign tissues. The positive rates and leves(x2 =43.16,x2Xk-w = 64. 19, P ＜0. 01)among the three groups were significantly different. When the critical value of Rac1 mRNA level was 1.73 × 104 copies/μl determined by ROC, the sensitivity and specificity were 88. 5% and 100% respectively. When the critical value of Rac1 level was 7.49 × 105 copies/μl, the sensitivity and specificity were 57.9% and 78. 6% respectively. ConclusionThe Rac1 mRNA level detected with fluorescence quantitative RT-PCR has reference value to distinguish between benign and malignant lesions and evaluate gastric carcinoma metastasis.