标题：蓝萼乙素对两种宫颈癌细胞裸鼠移植瘤的抑制作用及其机制

      作者：千文君，潘莹，白洁宇，何全中，孙力    (新乡医学院第三临床学院，河南 新乡 453000)

      卷次： 2017年28卷14期

      【摘要】 目的 探讨蓝萼乙素(GLB)对两种宫颈癌细胞株HeLa、SiHa裸鼠移植瘤的抑制作用及其机制。方法 将HeLa、SiHa细胞分别种植至BALB/C nu裸小鼠右大腿根部背侧皮下，建立裸小鼠移植瘤模型；待移植瘤体积约为0.1 cm3时，于第 10天将HeLa组随机分为 4组，第 14天将SiHa组随机分为 4组，分别为生理盐水组、10%乙醇组、GLB低剂量组和高剂量组，每组 5只；经腹腔注射给药，HeLa组给予生理盐水 0.4 mL/只、10%乙醇 0.4 mL/只、GLB 0.15 μmol/g 0.4 mL/只和0.20 μmol/g 0.4 mL/只，隔日1次，共20 d；SiHa组给予生理盐水0.4 mL/只、10%乙醇0.4 mL/只、GLB 0.18 μmol/g 0.4 mL/只和0.24 μmol/g 0.4 mL/只，隔日1次，共20 d。腹腔注射给药的同时，隔天用标准体重秤称量体质量并做好记录，每隔2 d专人用游标卡尺精确测量移植瘤的最大长径和短径，至少测量两次取平均值。给药结束后处死裸鼠前，眼球取血，测生化指标和肿瘤标志物，观察各组荷瘤鼠生化指标和肿瘤标志物的变化；通过移植瘤生长抑制试验，组织HE染色，观察各组移植瘤的大体形态、组织形态学的变化；免疫组化法检测移植瘤组织中自噬相关蛋白PTEN、Beclin 1、LC3的表达变化，Western blot法进一步检验相关蛋白变化。结果 裸鼠处死后，HeLa、SiHa组中生理盐水组、10%乙醇组、GLB低剂量组和高剂量组生化指标比较差异均无统计学意义(P>0.05)；GLB低剂量组和高剂量组的肿瘤标志物较生理盐水组、10%乙醇组降低，差异均有统计学意义(P<0.05)；GLB低剂量组和高剂量组的移植瘤体积均明显小于生理盐水组和 10%乙醇组，差异均有统计学意义(P<0.05)；HE染色显示：GLB低剂量组和高剂量组移植瘤组织出现凋亡、坏死改变；HeLa和SiHa组心肝脾肺肾均未发现转移瘤细胞；免疫组化法、Western-blot法结果示：GLB低剂量组和高剂量组的肿瘤组织PTEN、Beclin 1及LC3蛋白表达水平较生理盐水组和 10%乙醇组明显升高，差异均具有统计学意义(P<0.05)。结论 GLB对人宫颈癌HeLa和SiHa细胞移植瘤的生长具有明显的抑制作用，其机制可能与上调PTEN、Beclin 1、LC3的表达，诱导细胞自噬凋亡有关。
      【关键词】 宫颈癌；蓝萼乙素；裸鼠；移植瘤；抑瘤率
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2017）14—2237—06

Inhibitory effect of GLB on two kinds of cervical cancer cell lines transplanted tumor in nude mice and itsmechanism.
QIAN Wen-jun, PAN Ying, BAI Jie-yu, HE Quan-zhong, SUN Li. The Third Affiliated Hospital of XinxiangMedical University, Xinxiang 453000, Henan, CHINA
【Abstract】 Objective To study the inhibitory effect of Glaucocalyxin B (GLB) on two kinds of cervical cancercell lines HeLa and SiHa transplanted tumor in nude mice and its mechanism. Methods Human cervical cancer HeLaand SiHa cells were transplanted into nude mice in right thigh dorsal subcutaneously to establish the transplanted tumormodel of HeLa and SiHa cell. When the transplanted tumor volume was about 0.1 cm3, HeLa cell were randomly dividedinto 4 groups at the tenth day. At the fourteenth day, SiHa group were randomly divided into 4 groups, namely normal sa-line group, 10% ethanol group, GLB low dose group and high dose group, with 5 mice in each group. Through intraperito-neal injection, HeLa group was injected with 0.4 mL physiological saline, 0.4 mL 10% ethanol, 0.4 mL GLB (0.15 μmol/g)and 0.4 mL GLB (0.20 μmol/g), once every other day for 20 days. SiHa group was injected with 0.4 mL physiological sa-line, 0.4 mL 10% ethanol, 0.4 mL GLB (0.18 μmol/g) and 0.4 mL GLB (0.24 μmol/g), once every other day for 20 days.At the same time as intraperitoneal injection, the maximum diameter and minor diameter of the transplanted tumor wereaccurately measured at least two times for average by a Vernier caliper every 2 days. Before the administration of thenude mice, the nude mice were sacrificed and the blood samples were taken from the naked eye to measure and observechanges of biochemical indexes and tumor markers. Through the tumor growth inhibition test and transplanted tumor tis-sue HE staining, transplanted tumor gross morphology and histomorphological changes of each group were observed.The expression of phosphatase and tensin homolog deleted on chromosome ten (PTEN), Beclin 1 and LC3 in transplant-ed tumor tissues was detected by immunohistochemistry, and the changes of related proteins were detected by West-ern-blot. Results There was no significant difference in biochemical indexes for physiological saline group, 10% etha-nol group, GLB low dose group and high dose group (P>0.05). The tumor markers of GLB low dose group and high·论 著·doi:10.3969/j.issn.1003-6350.2017.14.001基金项目：河南省教育厅资助项目(编号：14B320004)