雌、孕激素对宫颈腺癌Hela细胞荷瘤裸鼠的影响

Estrogen and Progesterone Effects on the Growth of Tumor in Xenograft Nude Mice of Cervical Adenocarcinoma Hela Cell

目的评价激素干预对裸鼠体内He La肿瘤生长的影响,为临床进行宫颈腺癌激素替代治疗提供基础实验依据。方法采用免疫组化方法检测体外培养的人宫颈腺癌He La细胞雌、孕激素受体的表达;实验分四组:E2组、P组、E2+P组及对照组,每组8只四周龄裸鼠,每只裸鼠双侧后肢背部皮下注射Hela细胞悬液(0.1 ml),24 h后分别予E2(1μg)、P(150μg)和E2+P(1μg+150μg)及实验组等体积溶剂(1%DMSO+10%羟丙基-β环糊精的生理盐水0.1ml)腹腔注射治疗,每天一次;继续按上述方法给药,观察各组肿瘤大小、荷瘤裸鼠体重及生存时间的差异。结果免疫组化检测发现体外培养的Hela细胞Er、Pr均呈强阳性表达;其中E2组中8号裸鼠组右侧背部接种失败,E2组只接种15个部位,其余各组均接种16个部位,一周后经病理证实成功建立Hela细胞荷瘤裸鼠模型,E2组、P组、E2+P组及对照组成瘤率分别是:100%(15/15)、81.25%(13/16)、100%(16/16)、87.5%(14/16)(P>0.05);实验裸鼠共用药治疗82天,E2组、P组、E2+P组及对照组的平均肿瘤体积分别是450.17 mm^3、107.96 mm^3、168.94 mm^3、105.89 mm^3,其中E2与对照组相比,肿瘤体积更大(P<0.05),各组裸鼠体重之间差异无统计学意义(P>0.05);E2组的1号鼠腹腔转移,转移瘤体积为171.5 mm^3,E2+P组的2号和7号鼠分别有头部及腹腔转移,转移瘤体积分别为1070 mm^3和600 mm^3,并且均死亡,E2+P转移瘤体积明显更大。结论通过细胞悬液皮下注射法成功建立Hela细胞荷瘤裸鼠模型;在荷瘤裸鼠体内,雌二醇促进肿瘤生长,加用孕激素后可能促进肿瘤细胞的早期转移。

Objective To evaluate the influence of hormone intervention on the xenograft nude mice, to provide the basic experimental basis for hormone replacement therapy for cervical adenocarcinoma patients. Methods The expression of estrogen and progesterone receptor in human cervical adenocarcinoma Hela cells in vitro was detected by immunohistochemical method. The experiment was divided into 4 groups:group E2,group P, E2 + P group and control group. Hela cell suspension liquid(0. 1 ml) was injected subcutaneously into the back of bilateral hind limbs of nude mice. After 24 hours,E2 ( 1 μg),P (150 μg) and E2 + P (1μg) and of experimental group ( 1% DMSO + 10% hydroxypropyl-beta cyclodextrin + aline) were injected intraperitoneally once a day. 3. Continuous treatment by E2 ,P and E2 + P and menstruum on different group seperately. Results Er and Pr expression in Hela cells was strongly positive expression in Hela cells cultured in vitro. 2. 84 week age nude mices were distribute into E2 group, P group, E2 4- P group and control group separately (n = 8 ). In E2 group, only 15 sites were inoculated, However, 16 sites were inoculated in other 3 groups. The model of xenograft nude mices of Hela cells was successfully established by pathology 1 week after inoculation. The tumour growth rate of each group was:87. 5%( 14/16), 100%(15/15 ),81.25%( 13/16), 100%(16/16),The difference was statistically significant (P < 0. 05 ). After 24 hours of tumor inoculation, each group of nude mice was treated by E2, P, E2 + P and menstruum intraperitoneal injection. After 82 days of treatment in xenograft nude mices, the mean tumor volume of E2 group and E2 + P group were 105. 89 mm3,450. 17 mm3,107. 96 mm^3 and 168. 94 mm3 respectively. The tumor volume of E2 group was larger than the control group with statistically significant difference ( P < 0 . 0 5 ), but there was nosignificant difference in body weight of nude mice in each group (P >0. 05). NO. 1 xenograft nude mice of E2 group observed abdominal metastases tumor with volume 171.5 mm^3. NO. 2 and NO. 7 xenograft nude mice from E2 + P group with head and abdominal cavity metastasis tumor with the volume 1070 mm3 and 600 mm^3 respectively, and both died on 81 days. The metastatic rate of E2 group and E2 + P groups was 12. 5%(1/8) and 25%(2/8) respectively. The metastatic tumor volume of E2 + P groups was much larger than E2 group. Conclusion The xenograft nude mice model of Hela cells is successfully established by subcutaneous injection of cell suspension. In the xenograft nude mices, the estradiol promoted the growth of the tumor, and the addition of progestin may promote the early metastasis of the tumor possibility.