Objective： To study the antitumor effects and associated mechanisms of extract of the Smilax china L. rhizome （SCR） on ovarian cancer cells. Methods： Ovarian cancer cells A2780 were treated with different concentr...Objective： To study the antitumor effects and associated mechanisms of extract of the Smilax china L. rhizome （SCR） on ovarian cancer cells. Methods： Ovarian cancer cells A2780 were treated with different concentrations of SCR extract （SCRE）, and compared with controls. Effects on cell growth were evaluated by cell counting kit-8 （CCK-8） assay; proliferation effects by EdU incorporation assay; cell cycle by propidium iodide staining; apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide; cellular distribution of nuclear factor- κ B （NF-κ B） by immunofluorescence; protein levels of NF- κB, caspase-3, poly-adenosine diphosphate （ADP）-ribose polymerase （PARP）, Bcl-2-associated X protein （Bax）, cellular inhibitor of apoptosis （clAP）-1, anti-X-linked inhibitor of apoptosis protein （XlAP）, B-cell lymphoma-extra large （BcI-XL）, B-cell lymphoma-2 （Bcl-2） and AKT by Western blotting; and effects of SCRE combined with cisplatin or adriamycin on A2780 cells by CCK-8 assay. Results： SCRE suppressed A2780 cell proliferation in a dose-dependent manner （P〈0.05, /=〈0.01）, arrested cells in GJM phase and induced apoptosis by activating caspase-3, PARP and Bax. SCRE treatment also correlated with inhibition of NF-κ B and downregulation of Bcl-2, Bcl-XL, clAP-l, XlAP and AKT. SCRE can promote chemosensitivity to cisplatin and adriamycin in A2780 cells （P〈0.01）. Conclusion： SCR effectively inhibits NF- κ B, induces apoptosis and reduces chemoresistance to cisplatin and adriamycin in ovarian cancer cells, which might be its molecular basis for treating ovarian cancer.展开更多
目的:检测hTERT基因反义核酸对A2780细胞端粒酶活性的影响及其机制。方法:TRAP法检测端粒酶活性,流式细胞仪检测hTERT蛋白表达。结果:检测端粒酶活性,A2780细胞吸光度A值为(0.492±0.051),hTERT反义核酸作用48 h A值降为(0.351±...目的:检测hTERT基因反义核酸对A2780细胞端粒酶活性的影响及其机制。方法:TRAP法检测端粒酶活性,流式细胞仪检测hTERT蛋白表达。结果:检测端粒酶活性,A2780细胞吸光度A值为(0.492±0.051),hTERT反义核酸作用48 h A值降为(0.351±0.051),hTERT反义核酸作用72 h A值降为(0.238±0.024)检测hTERT蛋白表达,A2780细胞hTERT蛋白阳性率(89.513±3.389)%,hTERT反义核酸作用48hhTERT蛋白阳性率低至77.237±2.872%,hTERT反义核酸作用72 h hTERT蛋白阳性率低至(47.767±1.226)%。而hTERT正义核酸无上述作用。hTERT反义核酸作用A2780细胞48 h、72h对hTERTmRNA表达无影响。结论:hTERT反义核酸降低A2780细胞端粒酶活性,机制可能是降低hTERT蛋白表达量。展开更多
基金Partially supported by the Natural Science Foundation of Hubei Province(No.2013CFB389)the National Natural Science Foundation of China(No.81202962)
文摘Objective： To study the antitumor effects and associated mechanisms of extract of the Smilax china L. rhizome （SCR） on ovarian cancer cells. Methods： Ovarian cancer cells A2780 were treated with different concentrations of SCR extract （SCRE）, and compared with controls. Effects on cell growth were evaluated by cell counting kit-8 （CCK-8） assay; proliferation effects by EdU incorporation assay; cell cycle by propidium iodide staining; apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide; cellular distribution of nuclear factor- κ B （NF-κ B） by immunofluorescence; protein levels of NF- κB, caspase-3, poly-adenosine diphosphate （ADP）-ribose polymerase （PARP）, Bcl-2-associated X protein （Bax）, cellular inhibitor of apoptosis （clAP）-1, anti-X-linked inhibitor of apoptosis protein （XlAP）, B-cell lymphoma-extra large （BcI-XL）, B-cell lymphoma-2 （Bcl-2） and AKT by Western blotting; and effects of SCRE combined with cisplatin or adriamycin on A2780 cells by CCK-8 assay. Results： SCRE suppressed A2780 cell proliferation in a dose-dependent manner （P〈0.05, /=〈0.01）, arrested cells in GJM phase and induced apoptosis by activating caspase-3, PARP and Bax. SCRE treatment also correlated with inhibition of NF-κ B and downregulation of Bcl-2, Bcl-XL, clAP-l, XlAP and AKT. SCRE can promote chemosensitivity to cisplatin and adriamycin in A2780 cells （P〈0.01）. Conclusion： SCR effectively inhibits NF- κ B, induces apoptosis and reduces chemoresistance to cisplatin and adriamycin in ovarian cancer cells, which might be its molecular basis for treating ovarian cancer.
文摘目的:检测hTERT基因反义核酸对A2780细胞端粒酶活性的影响及其机制。方法:TRAP法检测端粒酶活性,流式细胞仪检测hTERT蛋白表达。结果:检测端粒酶活性,A2780细胞吸光度A值为(0.492±0.051),hTERT反义核酸作用48 h A值降为(0.351±0.051),hTERT反义核酸作用72 h A值降为(0.238±0.024)检测hTERT蛋白表达,A2780细胞hTERT蛋白阳性率(89.513±3.389)%,hTERT反义核酸作用48hhTERT蛋白阳性率低至77.237±2.872%,hTERT反义核酸作用72 h hTERT蛋白阳性率低至(47.767±1.226)%。而hTERT正义核酸无上述作用。hTERT反义核酸作用A2780细胞48 h、72h对hTERTmRNA表达无影响。结论:hTERT反义核酸降低A2780细胞端粒酶活性,机制可能是降低hTERT蛋白表达量。
