IL-15基因修饰的NK细胞对原代卵巢癌细胞的杀伤作用

Cytotoxic activity of IL-15 gene modified-NK cells against primary ovarian cancer cells

目的:探讨γ射线照射后的IL-15基因修饰的NK细胞(简称NK-ustc细胞)对原代卵巢癌细胞的体内外杀伤活性。方法:分离卵巢癌患者腹水原代卵巢癌细胞。不同剂量γ射线(0、1、2、4、8、16 Gy)照射NK-ustc细胞,3H-TdR掺入法检测照射后NK-ustc细胞的增殖情况,51Cr释放法检测NK-ustc细胞对K562和原代卵巢癌细胞的杀伤活性。建立人裸鼠荷卵巢癌模型,随机分为NK-ustc治疗组(模型鼠腹腔注射辐照后的NK-ustc细胞)和培养基对照组,同时设空白对照组(正常裸鼠腹腔注射辐照后的NK-ustc细胞),观察各组裸鼠体重、腹围及生存期。结果:1、2、4、8、16 Gy辐照后NK-ustc细胞的增殖率分别为(62.1±9.8)%、(41.3±8.7)%、(14.6±4.1)%、(0.1±0.03)%和(0.2±0.04)%。当效靶比为10∶1时,0、8 Gy辐照后NK-ustc细胞对K562的杀伤率分别为(45.4±8.9)%和(43.1±6.4)%,对原代卵巢癌细胞的杀伤率分别为(54.6±6.4)%和(48.3±5.8)%,说明辐照不影响NK-ustc细胞的杀伤活性(P>0.05)。辐照后NK-ustc细胞治疗组荷瘤小鼠的中位生存期为75 d,对照组为39 d(P<0.05),空白对照组小鼠全部存活。结论:γ射线照射可有效抑制NK-ustc细胞增殖,但保留该细胞对原代卵巢癌细胞的杀伤活性。

Objective： To explore the cytotoxic activity of IL-15 gene modified-NK cells （NK-ustc cells） against pri- mary ovarian cancer cells in vitro and in vivo. Methods： Primary ovarian cancer cells were isolated from ascites of patients. NK-uste cells were irradiated with different dosages of gamma ray （0, 1, 2, 4, 8, 16 Gy） , and the proliferation of irradiated NK-ustc cells were detected by 3 H-TdR incorporation assay. Cytotoxic activities of NK-ustc cells against K562 and primary ovarian cancer cells were measured by 51Cr release assay. The tumor-bearing mouse model was established using primary ovarian cancer cells and randomly divided into NK-ustc treatment group （ intraperitoneal injection of 8 Gy irradiated NK-ustc cells） and medium control group; moreover, blank control group （8 Gy irradiated NK-ustc cells were in- jected into nude mice） was also included in the present study. The body weight, abdomen circumference and survival time of nude mice were monitored. Results： After 1, 2, 4, 8 and 16 Gy irradiation, the proliferation rates of NK-ustc cells were （62. 1±9.8）% , （41.3 ±8.7）% , （14.6 ±4.1）%, （0.1±0.03）% and （0.2 ±0.04）% , respectively. The cytotoxic rates of 0 and 8 Gy irradiated-NK-ustc cells against K562 cells were （45.4 ± 8.9） % and （43.1 ±6.4） % when the effector to target ratio was 10： 1, and those against ovarian cancer cells were （54.6 ± 6.4）% and （48.3 ± 5.8 ）%, respectively. Thus, irradiation had no influence on cytotoxicity of NK-uste cells （P 〉 0.05 ）. The median survival time of irradiated-NK-ustc cells treated mice was 75 d, and that of control group was 39 d （P 〈 0.05 ）. All the mice in the blank control group survived. Conclusion： Gamma ray irradiation can effectively inhibit proliferation of NK-ustc cells, but retain their cytotoxic activities against primary ovarian cancer cells.