Elevated miR-27b-3p enhances the radioresistance by targeting PLK2 in breast cancer / 中华放射医学与防护杂志

Objective:To investigate the effect of miR-27b-3p on radiation resistance of breast cancer cells.Methods:The relative expression levels of miR-27b-3p in normal tissues and breast cancer tissues were analyzed through GEO database and verified by the qRT-PCR assay. Cloning formation, immunofluorescence, and EDU assay were used to assess the functions of miR-27b-3p on radioresistance of breast cancer cells. The luciferase reporter assay was used to verify whether miR-27b-3p directly targeted PLK2 mRNA. A rescue experiment was performed to identify the influence of PLK2 overexpression on miR-27b-3p regulated radioresistance.Results:The expression level of miR-27b-3p in both breast cancer tissues ( t=2.99, P<0.01) and breast cancer cells ( t=21.21, 32.88, P<0.05) was significantly higher than those in normal breast tissues and cells, especially, it was elevated in radioresistant MCF-7R cells ( t=25.63, P<0.05). Overexpression of miR-27b-3p enhanced the cloning efficiency of MCF-7 cells ( t=10.32, P<0.05), and had a protective effect on the proliferation of irradiated MCF-7 cells ( t=8.77, 8.26, 8.03, P<0.05). But interference of miR-27b-3p reduced the cloning efficiency and proliferation of MCF-7R cells ( t=40.00, P<0.05) after irradiation with different doses( t=8.54, 8.32, 8.23, P<0.05). Moreover, PLK2 was verified to be a direct target of miR-27b-3p, and overexpression of PLK2 inhibited miR-27b-3p-mediated radioresistance of breast cancer cells (MCF-7: t=9.66, P<0.05; MCF-7R: t=6.42, P<0.05). Conclusions:miR-27b-3p contributes to the radioresistance of breast cancer cells by targeting PLK2.

Mature cystic teratoma complicated with primary ovarian carcinoid:A report of 3 cases and literature review / 吉林大学学报(医学版)

Objective:To discuss the clinical characteristics,diagnosis,treatment experience and prognosis of mature cystic teratoma complicated with primary ovarian carcinoid,and to summarize its general charateriseics and to enhance the knowledge of clinician.Methods:Three female patients presented unilateral ovarian tumor in physical examination and were admitted to hospital.The tumor markers of three patients were normal. The preoperative diagnosis was ovarian tumor.Unilateral salpingo-oophorectomy was conducted in two patients.The resection of ovarian tumor was conducted in one patient.Results:The three patients underwent operation successfully.The postoperative pathological diagnosis was mature cystic teratoma complicated with primary ovarian carcinoid.The patients recovered well,who accepted 15-month follow-up with normal life and stable condition,and had no recurrence and metastasis.Conclusion:Mature cystic teratoma complicated with primary ovarian carcinoid is easy to be misdiagnosed before operation.Salpingo-oophorectomy is commonly used as the treatment method of mature cystic teratoma complicated with primary ovarian carcinoid and it can result in good effectiveness and better prognosis.

Identification and characterization of stem cells in an ovarian cancer cell line and examination their drug resistance / 中华妇产科杂志

Objective To isolate side population (SP) cells from an established ovarian cancer (OC)cell line,characterize these cells,and examine their drug resistance. Methods SP and non-SP (NSP) cells were isolated by fluorescence-activated cell sorting (FACS),and cultured in differential conditions,then detected their SP ratio to compare their capability of differentiation and self-renewal. Moreover,SP and NSP cell tumorigenesis was examined by subcutaneous and intraperitoneal injection of these cells to nonobes ediabetic(NOD)-severe combined immundeficient(SCID)mice. Drug resistance to cisplatin was examined by cell counting kit-8 (CCK-8).Results SP cells could be isolated stablly and insistently. There was(4.81 ± 0.43)%of SP cells in the established OC cell line and(4.89 ± 0.33)%of SP cells after cultured the isolated SP cells in differentiation condition,and there was no significant different between these two quantities (P>0.05). However,after cultured the NSP cells,there was only (0.10 ± 0.03)%of SP cells which was significantly lower than that contained in the OC cell line(P<0.01). In the tumorigenesis assay 1.0 × 103 SP cells were injected subcutaneously and formed the xenografted tumors in 6 weeks(3/3),and 1.0×104 NSP cells were injected subcutaneously and did not form xenografted tumors in 12 weeks(0). The tumorigenic capability of SP cells was higher than that of NSP cells(P<0.01). Both the original and the xenografted tumors were low differentiated serous cystadenocarcinomas and expressed the ovarian serous cystadenocarcinomas CA125 marker after stained by HE and immunohistochemistry. Simultaneously,the SP cells were also capable to form tumors as shown by intraperitoneal injection. In the drug resistance assay shown that the 50% inhibitory concentration (IC50) of the SP and NSP cells were respectively(2.33 ± 0.14)μg/ml and(1.60 ± 0.04)μg/ml(P<0.05). After treated the unsorted OC cells with cisplatin,the quantity of SP cells increased to(40.10 ± 4.22)%and there was significant difference,when compared to the untreated cells which was(4.81±0.43)%(P<0.01). The SP cells survival rate was(58.7± 3.3)%when treated with cisplatin at its IC50 dose,and the rate decreased to(7.2±1.3)%(P<0.01)when verapamil was present. Conclusions The SP cells could be isolated from the established OC cell line. They had the capacities of self-renewal,differentiation,and tumorigenesis,and the new tumor demonstrated the original tumor′s phenotype. The SP cells also had stem cells′ biological characteristics and is resistant to cisplatin.

Study of the invasiveness and tumour formation of Bcrp1 + HeLa cervical cancer cells / 中华妇产科杂志

Objective To make sure whether or not Bcrp1 is the marker of cervical cancer stem cells or not by studying the invasive ability and formation of tumors of Bcrp1 + phenotype HeLa cells.Methods The tumor cell migration and invasion assay were used by boyden chamber to identify the invasive ability of Bcrp1 + phenotype HeLa cells.The formation of tumors in vivo experiments were completed,in which the two groups of cells with different concentrations were inoculated in non obese diabetes-severe combined immunodeficiency disease ( NOD/SCID ) mice ( 1 × 104,1 × 105,1 × 106/ml ) and the differences of time,rate and volume in the formation of tumors between two groups were observed.Results ( 1 ) In the invasion assay,the amount of cells that invaded through the artificial basement membrane in Bcrp1 + group were 99 ± 14,which was significantly greater than those in Bcrp1- group ( 57 ± 13,P ＜ 0.05 ) ; the length of the Bcrp1 + group was ( 366 ± 52 ) μm,which was significantly greater than the Bcrp1 - group ( 301 ± 54) μm ( P ＜ 0.05 ).( 2 ) Following transplantation of 1 × 104 cells,only the Bcrp1 + cells formed tumors in NOD/SCID mice.When 1 × 105 or 1 × 106 cells were transplanted,the tumor incidence and the tumor mass were greater in the Bcrp1 + groups than those in the Bcrp1 - groups ( P ＜ 0.05 ).Conclusion Bcrp1 + HeLa cell have the greater capacity of invasive and the tumorigenicity,which may contain cancer stem cells.

RNF152, a novel lysosome localized E3 ligase with pro-apoptotic activities

RING finger protein 152 (RNF152) is a novel RING finger protein and has not been well characterized. We report here that RNF152 is a canonical RING finger protein and has E3 ligase activity. It is polyubiqitinated partly through Lys-48-linked ubiquitin chains in vivo and this phenomenon is dependent on its RING finger domain and transmembrane domain. RNF152 is localized in lysosomes and co-localized with LAMP3, a lysosome marker. Moreover, over-expression of RNF152 in Hela cells induces apoptosis. These results suggest that RNF152 is a lysosome localized E3 ligase with pro-apoptotic activities. It is the first E3 ligase identified so far that is involved in lysosome-related apoptosis.

Research of the characterization of Bcrp1+ HeLa cells / 中华妇产科杂志

Objective To make sure whether Bcrp1 is the marker of cervical cancer stem-like cells or not by studying the characterization of Bcrp1+ HeLa cells.Methods Immunofluorescence stained flow cytometry and electron microacope were used to sort and observe uhrastructures of Bctp1+ and Bcrp1- HeLa cells.Flow cytometry wag used to identify the cycle and the rate of apoptosis with annexin V in two group cells.The expression of proliferating cell nuclear antigen (PCNA)and caspase-3 were tested using western blot methed.Results (1)There were 7.1% Bcrp1+ cells and 92.9% Bcrol- cells in HeLa cells.Bcrp1+ HeLa cells were large in size of nuclear and nucleoli are clear.and there were rich of cytomicrosome and rough endoplasmic reticulum.After sorted and cultured for 24,48,72 hours,the adhesion in Bcrp1+ cells were 72.8%,81.1%,80.4%,respectively.While,they were 3.3%,18.7%,12.6%at each time for Bcrpl- cells(all P<0. 05 ). (2) There are more S phase cells in Bcrp1+ cells than that in Bcrp1- cells (54. 1% vs 21.1%, P <0. 05) ,while the percentage of G0/G1 and G2/M in Bcrp1 - cells were highter than those in Bcrp1 + cells (53.0% vs 44. 4% ,25.9% vs 1.5% ; all P <0. 05 ). The rate of apoptosis in Bcrp1+ cells was lower than that in Bcrp1 - cells (0. 2% vs 5.3%, P < 0. 05 ). ( 3 ) The expression of PCNA in Bcrp1 + cells was higher than that in Bcrp1- cells (3140 vs 2255, P< 0. 05 ), while the expression of caspase-3 of Bcrpl + cells was lower than that in Bcrp1 - cells ( 1970 vs 3551, P < 0. 05 ). Conclusion There are more vigor and ability of proliferation and lower rate of apeptosis in Bcrp1 + HeLa cells than those in Bcrp1 - cells ,which may be some characters of cervical cancer stem cells.

Isolation and detection of CK17~+,CD44v5~+ and Bcrp1~+ cells in HeLa cell line of cervical carinoma / 吉林大学学报(医学版)

Objective To identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells,and investigate the relationship between them and select cervical cancer stem cell markers.Methods Flow cytometry was used to identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells;after isolation,the expressions of CK17 and CD44v5 in Bcrp1+ and Bcrp1-phenotype HeLa cells were measured by immunocytochemistry.Results HeLa cells contained 11% Bcrp1+ cells,0.7% CK17+ cells and 0.1%CD44v5+ cells.Bcrp1+ HeLa cells contained CK17+ and CD44v5+ HeLa cells,but the Bcrp1-Hela cells did not(P

Expressions of CK17 and P63 in human cervical cancer tissues and significances / 吉林大学学报(医学版)

Objective To investigate the expressions of CK17 and P63 in cervical cancer tissues,and explore the possibility of CK17 and P63 as markers of cervical cancer stem cells.Methods The expressions of CK17 and P63 in cervical squamous cancer tissues(n=55) and normal cervix tissues(n=20) were detected with immunohistochemical method.Results ①CK17 expressed in reserve cell cytoplasm in normal cervical specimens.In the cervical cancer specimens,the expression of CK17 was increased and it seemed that CK17 was stained in the margin of the cancer nest and the tumor embolus.In specimens with high grade malignant cancer or after the chemotherapy,the expressions of CK17 were increased(P