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OBJECTIVE@#To explore whether casticin (CAS) suppresses stemness in cancer stem-like cells (CSLCs) obtained from human cervical cancer (CCSLCs) and the underlying mechanism.@*METHODS@#Spheres from HeLa and CaSki cells were used as CCSLCs. DNA methyltransferase 1 (DNMT1) activity and mRNA levels, self-renewal capability (Nanog and Sox2), and cancer stem cell markers (CD133 and CD44), were detected by a colorimetric DNMT activity/inhibition assay kit, quantitative real-time reverse transcription-polymerase chain reaction, sphere and colony formation assays, and immunoblot, respectively. Knockdown and overexpression of DNMT1 by transfection with shRNA and cDNA, respectively, were performed to explore the mechanism for action of CAS (0, 10, 30, and 100 nmol/L).@*RESULTS@#DNMT1 activity was increased in CCSLCs compared with HeLa and CaSki cells (P<0.05). In addition, HeLa-derived CCSLCs transfected with DNMT1 shRNA showed reduced sphere and colony formation abilities, and lower CD133, CD44, Nanog and Sox2 protein expressions (P<0.05). Conversely, overexpression of DNMT1 in HeLa cells exhibited the oppositive effects. Furthermore, CAS significantly reduced DNMT1 activity and transcription levels as well as stemness in HeLa-derived CCSLCs (P<0.05). Interestingly, DNMT1 knockdown enhanced the inhibitory effect of CAS on stemness. As expected, DNMT1 overexpression reversed the inhibitory effect of CAS on stemness in HeLa cells.@*CONCLUSION@#CAS effectively inhibits stemness in CCSLCs through suppression of DNMT1 activation, suggesting that CAS acts as a promising preventive and therapeutic candidate in cervical cancer.
Subject(s)
Female , Humans , Cell Line, Tumor , HeLa Cells , Neoplastic Stem Cells/metabolism , RNA, Small Interfering/metabolism , Uterine Cervical Neoplasms/metabolismABSTRACT
Objective To evaluate the relationship between clinicopathological features and central lymph nodes metastasis in patients with papillary thyroid microcarcinoma (PTMC),and aim to provide an appropriate operation in clinic.Methods The clinical data of 536 patients undergoing surgical treatment in Department of Thyroid Surgery,the First Affiliated Hospital of Zhengzhou University from January 2015 to December 2017,who had pathologically confirmed popillary thyroid microcarcinoma (PTMC) were retrospectively analyzed.Thyroidectomy and isthmectomy with central compartment neck dissection were performed in all patients.The patients were divided into two groups according to whether there existed central lymph node metastasis,and compared the differences of gender,age,number of cancer lesions,tumor breakthrough envelope,BRAFV600E gene mutation,and tumor maximum diameter in the central lymph node metastasis,respectively,by t-test x2-test.Univariate logistic regression analysis and multivariate binary logistic regression analysis were conducted to find risky factors.Results There existed difference between two groups by completing the gender,age,the BRAFV600E gene mutation and maximal tumor diameter(P ＜ 0.05).Univariate logistic regression analysis indicated that gender (P =0.046),age (P ＜ 0.01),maximal tumor diameter(P ＜0.01) and the BRAFV600E gene mutation(P =0.016) were significant predictors for central lymph nodes metastasis.And multivariate binary logistic regression analysis revealed that the rate of lymph node metastasis significantly increased in cases of larger tumor diameter(P ＜0.01),BRAFV600E gene mutation(P =0.035) and ageing below 45 years old (P ＜ 0.01).Conclusions The treatment for central lymph node metastasis of PTMC should be different considering elements including BRAF600E gene mutation prophylactic,ageing below 45 years old and larger tumor diameter.Therefore central lymphadenectomy should be performed when the primary lesion was resected.
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UPLC-QTOF-MS/MS was used to identify metabolites in rat blood, urine and feces after the administration of n-butanol extract derived from steamed notoginseng. The metabolic process of saponins came from steamed notoginseng was analyzed. The metabolites were processed by PeakView software, and identified according to the structural characteristics of prototype compounds and the accurate qualitative and quantitative changes of common metabolic pathways. Four saponins metabolites were identified based on MS/MS information of metabolites, namely ginsenoside Rh₄, Rk₃, Rk₁, Rg₅,and their 15 metabolites were verified. The metabolic pathways of the four ginsenosides in n-butanol extract included glucuronidation, desugar, sulfation, dehydromethylation, and branch loss. The metabolites of main active saponin components derived from steamed Panax notoginseng were analyzed from the perspective of qualitative analysis. And the material basis for the efficacy of steamed notoginseng was further clarified.