自然杀伤细胞系NK-92治疗卵巢癌的体外及动物实验研究

目的研究放射线照射后的自然杀伤细胞系NK92对人卵巢癌细胞的体外杀伤活性及对卵巢癌动物模型的治疗效果。方法医用电子直线加速器照射NK92细胞,照射剂量分别为0、1、2、4、8、16Gy,将NK92细胞按照上述不同照射剂量分为6组。台盼蓝拒染法进行细胞计数,氚标记的胸腺嘧啶核苷掺入法检测细胞增殖活性,51Cr释放法检测NK92细胞对靶细胞HO8910的杀伤活性。观察照射后的NK92细胞对NOD/SCID鼠卵巢癌皮下移植瘤的治疗效果,NOD/SCID鼠分肿瘤组(仅注射人卵巢癌细胞系HO8910细胞)和治疗组(同时注射HO8910和8Gy组NK92细胞)。结果(1)体外实验:4、8Gy组NK92细胞数量在照射后的第5天分别减少到10%、0%;照射后48h,与0Gy组NK92细胞相比,4、8Gy组增殖活性分别降至29%、6%。对HO8910细胞杀伤结果显示,4、8Gy组杀伤率分别在42%～62%、33%～58%之间。(2)动物实验:接种后30、40、50d,肿瘤组NOD/SCID鼠瘤结节体积分别为(0.214±0.061)cm3、(0.382±0.051)cm3、(0.428±0.021)cm3,治疗组分别为(0.047±0.019)cm3、(0.167±0.021)cm3、(0.343±0.022)cm3,治疗组与肿瘤组相比均有减小,两组比较,差异有统计学意义(P<0.01)。治疗组NOD/SCID鼠全部存活到120d,肿瘤组于接种后74～82d全部死亡,中位生存时间为76d,与治疗组比较,差异有统计学意义(P<0.01)。结论放射线照射后的NK92细胞治疗卵巢癌是有效的。

Effect of natural killer cell line NK-92 against human ovarian carcinoma cells in vitro and in vivo

OBJECTIVE: To study the cytotoxic activity of NK-92 cells irradiated against human ovarian cancer. METHODS: NK-92 cells were exposed to different doses of radiation and assayed for proliferation by a standard (3)H-thymidine incorporation assay and cell count by using trypan blue exclusion. The cytotoxic activity of NK-92 cells against targets was measured in a standard (51)Cr-release assay in vitro. The effectiveness of irradiated NK-92 cells on ovarian cancer was compared with the control group of cancers (without injection of irradiated NK-92 cells). RESULTS: (1) In vitro:The proliferation of NK-92 cells was inhibited by radiation of 4, 8 and 16 Gy, respectively. From the (3)H-thymidine incorporation data, irradiation by 4 Gy reduced cell proliferation to 29% of control, while 8 Gy reduced proliferation to 6%. The cytotoxicity of NK-92 cells at 4 Gy 2 days following irradiation was approximately 42%-62% for ovarian cancer cell HO-8910, while it was 33%-58% at 8 Gy. (2) In vivo: Tumor size in treatment group was (0.047 +/- 0.019) cm(3) on day 30 after inoculation, and (0.167 +/- 0.021) cm(3) on day 40 and (0.343 +/- 0.022) cm(3) on day 50, while the sizes were smaller in treatment group (P < 0.01). In addition, the tumor group animals died between 74-82 days after injection of HO-8910 cells, while the treatment group animals were alive over 120 days (P < 0.01). CONCLUSION: Our study indicates that injection of irradiated NK-92 cells may be a potentially effective treatment for human ovarian carcinoma.