1.8-bromo-7-methoxychrysin reverses EMT of the sphere-forming cells derived from cervical cancer SiHa cell line
Xiaozheng CAO ; Xiaohong YANG ; Jianguo CAO ; Honglin XIANG
Chinese Pharmacological Bulletin 2014;(5):668-671
Aim To investigate whether 8-bromo-7-methoxychrysin ( BrMC ) can reverse the epithelial-mesenchymal transition ( EMT) in sphere-forming cells (SFCs) derived from cervical cancer SiHa cell line. Methods Human cervical cancer SiHa cell line was cultured in vitro and then SFCs were obtained from Si-Ha cell line using suspension culture with the stem-condition culture system. After treatment with BrMC for 72 h, the morphology of SFCs of SiHa cell line was observed via monolayer adherence culture. The expres-sion levels of E-cadherin, N-cadherin and Twist1 pro-teins were analyzed using western blot assay. Results SFCs of SiHa cell line presented the mesenchyme-like( spindle-shaped) cell shape and exhibited reduced E-cadherin and elevated N-cadherin protein expres-sion, compared with the parental cells. After treatment with BrMC(1. 0 μmol·L-1 ) for 72 h, SFCs of SiHa cell line changed from spindle-shaped cells in to poly-gon shaped cells under monolayer adherence culture. After treatment with BrMC for 24 h, the expression lev-els of E-cadherin protein were up-regulated and the ex-pression levels of N-cadherin and Twist1 proteins were decreased in SFCs derived from SiHa cell line. Con-clusion BrMC effectively reverses EMT of SFCs de-rived from cervical cancer SiHa cell line, and its mechanism is involved in down-regulating the expres-sion of Twist protein.
2.Inhibition of PKD2 mediated apoptosis induced by 5, 7-dihydrox-8-nitrochrysin in U937 cell line
Jianling YANG ; Xiaozheng CAO ; Lihua HE ; Jianguo CAO ; Jianfeng YANG
Journal of Chinese Physician 2014;(6):728-730
Objective To investigate whether 5, 7-dihydrox-8-nitrochrysin (NOC) induces apoptosis in U937 monocytic leukae-mia cells is involved in the regulation of the activity of PKD 2.Methods U937 cell line cells were cultured in vitro .Apoptosis rate was analyzed by flow cytometry (FCM) using propidium iodide (PI) staining.DNA ladder bands were observed by DNA agarose gel electro-phoresis.The phosphorylated protein expression of PKD 2 was analyzed using Western blot .Results NOC (2.5, 5.0, and 10.0μmol/L) increased apoptosis rate in U937 cells in a concentration-dependent manner ( P <0.05).After treatment with NOC (5.0 and 10.0μmol/L) for 24 h, U937 cells presented typical DNA ladder bands .At the same time, not only did NOC effectively down-regulate the ex-pression of PDK2 phosphorylated protein , but also increased apoptosis rate in U 937 cells in the presence of G?6976, a specific inhibitor of PKD2.Conclusions The effect that NOC induces apoptosis in U 937 cells is related to the inhibition of the activity of PKD 2.
3.The effects of casticin on phosphorylation of FoxO3a in stem-like cells derived from ovarian cancer SKOV3 cell line
Ling JIANG ; Xiaozheng CAO ; Cheng LI ; Qiao XIAO ; Jianfeng YANG
China Oncology 2015;(5):360-364
Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.
4.Clinicopathologic characteristics and histogenesis of mucinous tumor of peritoneum.
Heping ZHANG ; Yu SUN ; Xiaozheng HUANG ; Ling JIA ; Yanhua BAI ; Dengfeng CAO
Chinese Journal of Pathology 2014;43(3):163-168
OBJECTIVETo investigate the clinicopathologic characteristics, prognosis and histologic origin of the mucinous tumor of the peritoneum.
METHODSAccording to 2010 WHO classification of tumours of the digestive system, 34 cases diagnosed as "pseudomyxoma peritonei (PMP) " were reevaluated and divided into low grade and high grade. Immunohistochemistry was applied to investigate the expression of SATB2 and the histologic origin of the mucinous tumor of the peritoneum, using antibodies against SATB2, CK7, CK20 and CDX-2. The relationship between clinicopathologic characteristics and prognosis of the low grade and high grade tumors were analyzed.
RESULTSTwenty five patients had low grade mucinous tumors (two of them were no cell type), nine patients had high grade mucinous tumors. There was no significant difference between low grade and high grade mucinous tumors in age, sex, recurrence and organs involvement (P>0.05). Thirty patients were followed up, the overall survival rates of patients with low grade and high grade mucinous tumors were 13/21 (61.9%) and 3/9, respectively. The median survival time was 74 and 24 months in low and high grade patients, and the difference was statistically significant (P=0.002).Immunohistochemistry showed the expression rates of CDX-2, CK20, and CK7 in totally 32 cases (excluding 2 cases of no cell type) were 30/32(93.8%), 31/32 (96.9%), and 3/16, respectively; the expression rates of CDX-2, CK20, and CK7 in 16 cases with distinct primary site were 15, 16, and 1, respectively; fifteen of 16 cases of tumors of unknown primary site were positive for CDX-2 and CK20, two of the them were positive for CK7. There was no difference in the expression of CDX-2, CK20 and CK7 between tumors with distinct primary site and tumors with unknown primary site (P>0.05). The expression rate of SATB2 in the cases was 56.3% (18/32), excluding 2 cases of no cell type. There was no significant difference between low grade and high grade tumors in the expression of SATB2 [15/23(65.2%) vs 3/9, P=0.102], also SATB2 was not related to the prognosis of the tumor (P=0.786).
CONCLUSIONThe prognosis of the mucinous tumor of the peritoneum was significantly different between low grade and high grade according to WHO 2010 classification, and most mucinous tumor of the peritoneum originated from the appendix.
Adenocarcinoma, Mucinous ; metabolism ; pathology ; secondary ; surgery ; Adult ; Aged ; Aged, 80 and over ; Appendiceal Neoplasms ; pathology ; surgery ; CDX2 Transcription Factor ; Female ; Follow-Up Studies ; Homeodomain Proteins ; metabolism ; Humans ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Lymphatic Metastasis ; Male ; Matrix Attachment Region Binding Proteins ; metabolism ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Peritoneal Neoplasms ; metabolism ; pathology ; secondary ; surgery ; Pseudomyxoma Peritonei ; metabolism ; pathology ; surgery ; Survival Rate ; Transcription Factors ; metabolism