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Objective:To explore the value of cell division cycle 42 (CDC42) in the efficacy and prognosis evaluation of arterial infusion chemotherapy for pancreatic cancer.Methods:The clinical data of 100 patients with pancreatic cancer who underwent arterial infusion chemotherapy from January 2018 to January 2020 at Second People's Hospital of Wuhu were retrospectively analyzed, and all patients were divided into effective group (the complete remission and partial remission) and ineffective group (the stable disease and the progressive disease) according to the chemotherapy efficacy determined by CT. The clinicopathological characteristics of both groups were compared. The influencing factors of chemotherapy efficacy were determined by using multivariate logistic regression model analysis. The efficacy evaluated by CT examination was treated as the gold standard. The receiver operating characteristic (ROC) curve was used to analyze the value of CDC42 level predicting the efficacy of arterial infusion chemotherapy for pancreatic cancer patients before infusion chemotherapy. Survival analysis was performed by using Kaplan-Meier and log-rank test was also performed.Results:Among 100 patients with pancreatic cancer, there were 13 cases of complete remission, 30 cases of partial remission, 20 cases of stable disease, 37 cases of progressive disease; 43 cases in effective group and 57 cases in ineffective group. The proportions of tumor long diameter > 4 cm, TNM staging Ⅲ-Ⅳ, carcinoembryonic antigen (CA)199 > 37 U/ml, carcino-embryonic antigen (CEA) > 5 ng/ml, neutrophil-to-lymphocyte (NLR) > 2.8, serum total bilirubin > 34.2 μmol/L before infusion, CDC42 ≤ 1.11 μg/L, low differentiation degree and vascular invasion in ineffective group were higher than those in effective group (all P < 0.05). Tumor long diameter > 4 cm, TNM staging Ⅲ-Ⅳ, CA199 > 37 U/ml, CEA > 5 ng/ml before infusion, low differentiation degree, vascular invasion, and CDC42 ≤ 1.11 μg/L were independent risk factors for effectiveness of arterial infusion chemotherapy (all P < 0.05). The area under the ROC curve of CDC42 predicting the ineffectiveness of arterial infusion chemotherapy was 0.810 (95% CI 0.781-0.839, P <0.01), and the optimal cut-off value was 1.11 μg/L, the sensitivity was 96.25%, and the specificity was 63.13%. Survival curve analysis showed that the 2-year overall survival rate of patients with CDC42 > 1.11 μg/L was 58.93% which was greater than that of patients with CDC42 ≤ 1.11 μg/L (22.73%), and the difference was statistically significant ( χ2 = 14.99, P<0.001). Conclusion:CDC42 level is an independent influencing factor for the efficacy of arterial infusion chemotherapy in patients with pancreatic cancer, and it can effectively predict the prognosis of patients.
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Objective To study the effect of CKS2 on filopodia formation of A2780 cells through regulating CDC42 alternative splicing. Methods The filopodia were observed after CKS2 of A2780 cells were knocked down by lentivirus-mediated shRNA; the migrating ability of A2780 cells was also measured by wound healing assay; the expression levels of splicing variants CDC42-V1 and CDC42-V2 mRNA was determined by real time PCR after CKS2 knockdown. The expression levels of CKS2, CDC42-V1 and CDC42-V2 mRNA were further measured in each ovarian cancer and normal ovarian samples in the same way. Results The filopodia on A2780 cells obviously decreased, and the migrating ability of A2780 cells was also remarkably reduced after CKS2 knockdown (P<0.05). Meanwhile the expression of CDC42-V1 mRNA decreased and CDC42-V2 mRNA increased after CKS2 knockdown (P<0.05). Real time PCR results showed that the mRNA expression levels of CKS2 and CDC42-V1 were higher in ovarian cancer samples than in normal ovarian tissues; however, the expression level of CDC42-V2 mRNA was lower in ovarian cancer samples than in normal ovarian tissues (P<0.05). ConclusionCKS2 may affect the filopodia formation of A2780 cells through regulating CDC42 alternative splicing, and further affect the migrating ability of A2780 cells.
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BACKGROUND:Tougu Xiaotong Capsule has pretty good clinical therapeutic effect on osteoarthritis of early and middle periods. However, the mechanism of Tougu Xiaotong Capsule is not ful y clarified. The RhoA GTPases can regulate chondrocyte apoptosis and hypertrophy. OBJECTIVE:To observe the Tougu Xiaotong Capsule on the expression of Rac1and Cdc42 in tumor necrosis factor-α-induced in vitro cultured rat articular chondrocytes, and to explore its mechanism of action for combating osteoarthritis. METHODS:Knee cartilage of the 4-week-old Sprague-Dawley rats was used to stably establish in vitro culture system of chondrocytes. Passage 3 chondrocytes were identified by toluidine blue staining. Chondrocyte apoptosis was successful y induced by 20μg/L tumor necrosis factor-αand then Tougu Xiaotong Capsule at different dosage (500, 100, 20 mg/L) was given after 24-hour incubation. MTT assay was used to detect cellsurvival, flow cytrometry to measure mitochondrial membrane potential, and western blot assay to determine the protein expression of Rac1, Cdc42, Bax and Bcl-2. RESULTS AND CONCLUSION:Tougu Xiaotong Capsule could reduce tumor necrosis factor-α-induced apoptosis of chondrocytes to improve the survival rate of the cells, and at the same time, could down-regulate the protein expression of Rac1, Cdc42 and Bax and increase the protein expression of Bcl-2 significantly (P<0.05). Tougu Xiaotong Capsule possibly plays a therapeutic efficacy on osteoarthritis by reducing promote apoptosis Rac1, Cdc42 and Bax expression and increasing apoptosis inhibiting gene Bcl-2 expression, thereby to inhibit apoptosis of chondrocytes.
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BACKGROUND:The homing ability of mesenchymal stem cells is closely associated with the effects of celltransplantation. Clarifying the mechanism of chemotaxis and migration wil contribute to enhance the clinical application of mesenchymal stem cells. OBJECTIVE:To investigate the effect of Cdc42 in the homing of human umbilical cord mesenchymal stem cells. METHODS:First, mesenchymal stem cells were isolated from human umbilical cord, and co-cultured with tumor necrosis factorα, interleukin-1β, and transforming growth factorβ. Western blot assay was used to test the level of Cdc42. Besides, Cdc42 siRNA was synthesized by chemical method to transfect the cells, and cellmigration and adhesion were measured by Transwel and Matrigel separately. Meanwhile, the activity of signal molecule, extracellular regulated protein kinase 1/2, was evaluated by western blot. RESULTS AND CONCLUSION:The results indicated that the inflammation factors induced the highly expression of Cdc42 in human umbilical cord mesenchymal stem cells, almost double level to controls. siRNA notably inhibited the migration and adhesion of human umbilical cord mesenchymal stem cells through Cdc42 down-regulation, and the extracellular regulated protein kinase 1/2 and phosphorylation form were also decreased simultaneously. In a word, we speculate Cdc42 plays a role in the chemotaxis of human umbilical cord mesenchymal stem cells in vitro.
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Objective: To investigate the effect of cell division cycle 42 (Cdc 42) gene on the formation of filopodia suppressed by non-steroidal anti-inflammatory drugs (NSAIDs), and to explore the probable signal transduction pathway involving Cdc 42 in the inhibition effects on migration and invasion abilities of breast cancer cells induced by NSAIDs. Methods: MCF-7 cells were divided into four groups: blank control group, NS-398 (100 μmol/L)-treated group, cyclooxygenase-2 (Cox-2) small interfering RNA (siRNA)-transfected group and the negative siRNA-transfected group. The expressions of Cox-2 and Cdc42 mRNAs were detected by real-time fluorogentic quantitative PCR(RFQ-PCR), and the expressions of Cox-2 and Cdc42 proteins were measured by Western blotting. Actin-tracker Green fluorescent probe was used to examined the morphology of filopodias in MCF-7 cells. The invasion ability of MCF-7 cells was detected by using the Martrigel-coated Transwell. Results: The filopodias in MCF-7 cells disappeared in the NS-398-treated group, and the invasion ability of MCF-7 cells was also significantly decreased in this group compared with that in the blank control group (P0.05), and the difference was also not seen in the expression of Cdc42 mRNA or protein between these two groups (P>0.05). The active-Cdc42 level was significantly decreased in the NS-398-treated group compared with that in the blank control group (P<0.05). The filopodias in the Cox-2 siRNA-transfected group disappeared, and the invasion ability of MCF-7 cells transfected with Cox-2 siRNA was significantly decreased compared with that in the negative siRNA-transfected group (P<0.05). The active-Cdc42 level was also significantly decreased in MCF-7 cells transfected with Cox-2 siRNA compared with those transfected with negative siRNA (P<0.05). Conclusion: NSAIDs can suppress the invasion ability of breast cancer cells. This effect may be achieved by inhibiting the activity of Cox-2, decreasing the expression level of active-Cdc42, and suppressing the formation of filopodias.
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Objective To detect the proteins levels of RhoA and CDC42 in bone marrow mononucleated cells (BMMC) of patients with primary acute leukemia,and further determine the role of abnormal interactions between hematopoietic progenitor and bone marrow microenvironment on abnormal behaviors of leukemia cells. Methods BMMC samples were separated from 54 primary acute leukemia patients and 22 normal donors and the cell lysis samples were prepared. RhoA and CDC42 proteins were determined by Western blotting. Independent pair T test was conducted to evaluate whether the differences in RhoA and CDC42 expression were statistically significant between leukemia patients and normal donors. Spearman was applied in analyzing the correlation between expression of RhoA and CDC42 proteins and clinical characters of patients. Results RhoA and CDC42 proteins level of primary acute leukemia patients was significantly higher than that of normal samples. Especially, patients with M2,M3 and M5 subtypes exhibited significant higher RhoA proteins levels and M3 subtype exhibited significant higher CDC42 protein levels. Conclusion RhoA and CDC42 protein levels of primary acute leukemia patients are significantly higher than that of normal donors. This result suggests that RhoA and CDC42 associated efficient migration of leukemia cells could be implicated in abnormal interaction of leukemic cell with bone marrow microenvironment.