中国肿瘤临床
中國腫瘤臨床
중국종류림상
CHINESE JOURNAL OF CLINICAL ONCOLOGY
2014年
13期
861-865
,共5页
李圃%李冬田%尹冰楠%汤华%尹利荣
李圃%李鼕田%尹冰楠%湯華%尹利榮
리포%리동전%윤빙남%탕화%윤리영
人卵巢癌细胞%胞嘧啶脱氨酶基因%内皮抑素%重组腺病毒%基因治疗
人卵巢癌細胞%胞嘧啶脫氨酶基因%內皮抑素%重組腺病毒%基因治療
인란소암세포%포밀정탈안매기인%내피억소%중조선병독%기인치료
human ovarian cancer%cytidine deaminase%endostadine%adenovirus%gene therapy
目的:构建含CDglyES融合基因的重组腺病毒,体外观察其对人卵巢癌细胞的直接抑制作用及其对血管内皮细胞生长的抑制作用。方法:用细菌内同源重组方法构建腺病毒穿梭质粒prAdCDglyES。采用脂质体介导转染293包装细胞,扩增获取重组腺病毒rAdCDglyES。采用MTT法观察rAdCDglyES对卵巢癌细胞SKOV-3生长抑制的影响,同时观察其表达产物抑制脐静脉血管内皮细胞ECV-304增殖的作用。结果:纯化的rAdCDglyES滴度是1×1013.3 TCID50/L,其对SKOV-3细胞生长抑制率是(83.1±6.3)%,与对照组rAd-LacZ(24.1±13.2)%相比有显著性差异(P<0.01)。浓缩的转染rAdCDglyES的细胞培养上清液抑制ECV-304细胞增殖,抑制率是(78.7±1.6)%,而同样浓缩的转染rAd-CD的细胞培养上清液抑制率是(23.9±9.7)%,二者有显著性差异(P<0.01)。结论:研究结果表明重组腺病毒rAdCDglyES具有体外直接和间接抑制人卵巢癌细胞效能。
目的:構建含CDglyES融閤基因的重組腺病毒,體外觀察其對人卵巢癌細胞的直接抑製作用及其對血管內皮細胞生長的抑製作用。方法:用細菌內同源重組方法構建腺病毒穿梭質粒prAdCDglyES。採用脂質體介導轉染293包裝細胞,擴增穫取重組腺病毒rAdCDglyES。採用MTT法觀察rAdCDglyES對卵巢癌細胞SKOV-3生長抑製的影響,同時觀察其錶達產物抑製臍靜脈血管內皮細胞ECV-304增殖的作用。結果:純化的rAdCDglyES滴度是1×1013.3 TCID50/L,其對SKOV-3細胞生長抑製率是(83.1±6.3)%,與對照組rAd-LacZ(24.1±13.2)%相比有顯著性差異(P<0.01)。濃縮的轉染rAdCDglyES的細胞培養上清液抑製ECV-304細胞增殖,抑製率是(78.7±1.6)%,而同樣濃縮的轉染rAd-CD的細胞培養上清液抑製率是(23.9±9.7)%,二者有顯著性差異(P<0.01)。結論:研究結果錶明重組腺病毒rAdCDglyES具有體外直接和間接抑製人卵巢癌細胞效能。
목적:구건함CDglyES융합기인적중조선병독,체외관찰기대인란소암세포적직접억제작용급기대혈관내피세포생장적억제작용。방법:용세균내동원중조방법구건선병독천사질립prAdCDglyES。채용지질체개도전염293포장세포,확증획취중조선병독rAdCDglyES。채용MTT법관찰rAdCDglyES대란소암세포SKOV-3생장억제적영향,동시관찰기표체산물억제제정맥혈관내피세포ECV-304증식적작용。결과:순화적rAdCDglyES적도시1×1013.3 TCID50/L,기대SKOV-3세포생장억제솔시(83.1±6.3)%,여대조조rAd-LacZ(24.1±13.2)%상비유현저성차이(P<0.01)。농축적전염rAdCDglyES적세포배양상청액억제ECV-304세포증식,억제솔시(78.7±1.6)%,이동양농축적전염rAd-CD적세포배양상청액억제솔시(23.9±9.7)%,이자유현저성차이(P<0.01)。결론:연구결과표명중조선병독rAdCDglyES구유체외직접화간접억제인란소암세포효능。
To construct a recombinant adenovirus containing CDglyES fusion gene, which can directly inhibit human ovarian cancer cell and indirectly inhibit vascular endothelial cell growth. Methods:We constructed prAdCDglyES using a homolo-gous recombination method in bacteria. The prAdCDglyES was transfected to 293 packaging cells using liposome, in which rAdCDgly-ES was packaged and amplified. MTT was used to observe the proliferation inhibition effect of rAdCDglyES on human ovarian cancer cells and the growth inhibition effect of expressing products of rAdCDglyES on ECV-304. Results:The titer of rAdCDglyES was 1 × 1013.3 TCID50/L, whereas the inhibition rate on human ovarian cancer cell SKOV-3 was (83.1±6.3)%. This result is significantly different from the control rAd-LacZ, which had an inhibition rate of (24.1 ± 13.2)% (P<0.01). The concentrated culture supernatant from cells transfected with rAdCDglyES can inhibit ECV-304 cell proliferation at a rate of (78.7 ± 1.6)%. This rate is significantly different com-pared with that of the control with the same concentration of culture supernatant from cells transfected with rAd-CD, with an effect on ECV-304 cell shown by an inhibition rate of (23.9 ± 9.7)%(P<0.01). Conclusion:The results showed that the recombinant adenovirus rAdCDglyES could inhibit human ovarian cancer cells directly and indirectly.