姜黄素对宫颈癌大鼠的影响及作用机制研究
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摘要
目的研究姜黄素对宫颈癌大鼠的影响及作用机制。方法用人宫颈癌HeLa细胞悬液在大鼠左侧腋窝处皮下注射建立宫颈癌模型;空白组予以0. 9%Na Cl左侧腋窝处皮下注射。按照体重将造模成功大鼠随机分为5组:模型组、对照组和低、中、高3个剂量实验组,每组13只。模型组和空白组均腹腔注射0. 9%NaCl 1. 25 mL·kg~(-1);对照组腹腔注射16 mg·m L~(-1)氟尿嘧啶溶液1. 25mL·kg~(-1);低、中、高3个剂量实验组分别腹腔注射48,96,192 mg·mL~(-1)姜黄素溶液1. 25 m L·kg~(-1),均隔日给药1次,连续20 d。用免疫组化法检测肿瘤组织微血管密度(MVD),用实时荧光定量聚合酶链反应检测肿瘤组织微小核糖核酸-21(miR-21)和信号转导及转录活化因子-3 (Stat-3)的基因表达水平。结果空白组、模型组、对照组和低、中、高3个剂量实验组的MVD分别为(6. 26±0. 72),(23. 18±3. 04),(10. 25±1. 21),(17. 15±2. 33),(13. 84±1. 58)和(10. 62±1. 36)个;上述6组的miR-21表达水平(2~(-ΔΔCt)值)分别为0. 44±0. 06,1. 13±0. 15,0. 63±0. 06,1. 03±0. 12,0. 87±0. 09和0. 64±0. 07;上述6组的Stat-3表达水平(2~(-ΔΔCt)值)分别为0. 19±0. 02,0. 67±0. 09,0. 26±0. 03,0. 54±0. 07,0. 42±0. 06和0. 28±0. 04;模型组与空白组相比,上述指标的差异均有统计学意义(均P <0. 05),对照组和低、中、高3个剂量实验组与模型组相比,上述指标的差异均有统计学意义(均P <0. 05)。结论姜黄素能明显抑制宫颈癌大鼠肿瘤生长,抑制肿瘤组织的miR-21、Stat-3基因的表达水平,且药物浓度越高作用效果越明显。
Objective To analyze the effect of curcumin on cervical cancer model rats and its mechanism. Methods The rats were randomly divided into blank group and model building group. The model building group was established cervical cancer model by subcutaneous injection of human cervical cancer HeLa cell suspension into the left axilla;rats in the blank group received subcutaneous injection of physiological saline into the left axilla. Successful model of rats were randomly divided into five groups: model group,control group and low,midddle and high doses experimental groups,with 13 rats in each group. The model group and blank group were given physiological saline solution 1. 25 mL·kg~(-1) by intraperitoneal injection,control group was given 16 mg·mL~(-1) fluorouracil solution 1. 25 mL·kg~(-1) by intraperitoneal injection,three doses experimental groups were given 48,96,192 mg·mL~(-1) curcumin solution1. 25 mL·kg~(-1) by intraperitoneal injection; 1 time every other day for 20 days. Microvessel density( MVD) of tumor tissue was detected by immunohistochemistry. The expression levels of micro ribonucleic acid-21( microRNA-21) mRNA,signal transducer and activator of transcription-3( Stat-3) mRNA in tumor tissues was detected by real-time fluorescence quantitative polymerase chain reaction. Results MVD in blank group,model group,control group and low,middle and high doses experimental groups were respectively( 6. 26 ± 0. 72),( 23. 18 ± 3. 04),( 10. 25 ± 1. 21),( 17. 15 ± 2. 33),( 13. 84 ± 1. 58),( 10. 62 ± 1. 36) microvessels; the expression level of miRNA-21( 2~(-ΔΔCt) values) in the above six groups were respectively 0. 44 ± 0. 06,1. 13 ± 0. 15,0. 63 ± 0. 06,1. 03 ± 0. 12,0. 87 ± 0. 09,0. 64 ± 0. 07; the expression level of Stat-3( 2~(-ΔΔCt) values) in the above six groups were respectively 0. 19 ± 0. 02,0. 67 ± 0. 09,0. 26 ± 0. 03,0. 54 ± 0. 07,0. 42 ± 0. 06,0. 28 ± 0. 04. Comparing between model group with blank group,the difference of the factors were significant( all P < 0. 05); comparing between control group and three doses experimental groups with model group,the difference of the factors were all significant( all P < 0. 05). Conclusion Curcumin can significantly inhi-bit the growth of cervical cancer in rats model,inhibit the expression of miR-21 and Stat-3 in tumor tissues,and the higher the drug dose is,the more obvious the effect is.
Objective To analyze the effect of curcumin on cervical cancer model rats and its mechanism. Methods The rats were randomly divided into blank group and model building group. The model building group was established cervical cancer model by subcutaneous injection of human cervical cancer HeLa cell suspension into the left axilla;rats in the blank group received subcutaneous injection of physiological saline into the left axilla. Successful model of rats were randomly divided into five groups: model group,control group and low,midddle and high doses experimental groups,with 13 rats in each group. The model group and blank group were given physiological saline solution 1. 25 mL·kg~(-1) by intraperitoneal injection,control group was given 16 mg·mL~(-1) fluorouracil solution 1. 25 mL·kg~(-1) by intraperitoneal injection,three doses experimental groups were given 48,96,192 mg·mL~(-1) curcumin solution1. 25 mL·kg~(-1) by intraperitoneal injection; 1 time every other day for 20 days. Microvessel density( MVD) of tumor tissue was detected by immunohistochemistry. The expression levels of micro ribonucleic acid-21( microRNA-21) mRNA,signal transducer and activator of transcription-3( Stat-3) mRNA in tumor tissues was detected by real-time fluorescence quantitative polymerase chain reaction. Results MVD in blank group,model group,control group and low,middle and high doses experimental groups were respectively( 6. 26 ± 0. 72),( 23. 18 ± 3. 04),( 10. 25 ± 1. 21),( 17. 15 ± 2. 33),( 13. 84 ± 1. 58),( 10. 62 ± 1. 36) microvessels; the expression level of miRNA-21( 2~(-ΔΔCt) values) in the above six groups were respectively 0. 44 ± 0. 06,1. 13 ± 0. 15,0. 63 ± 0. 06,1. 03 ± 0. 12,0. 87 ± 0. 09,0. 64 ± 0. 07; the expression level of Stat-3( 2~(-ΔΔCt) values) in the above six groups were respectively 0. 19 ± 0. 02,0. 67 ± 0. 09,0. 26 ± 0. 03,0. 54 ± 0. 07,0. 42 ± 0. 06,0. 28 ± 0. 04. Comparing between model group with blank group,the difference of the factors were significant( all P < 0. 05); comparing between control group and three doses experimental groups with model group,the difference of the factors were all significant( all P < 0. 05). Conclusion Curcumin can significantly inhi-bit the growth of cervical cancer in rats model,inhibit the expression of miR-21 and Stat-3 in tumor tissues,and the higher the drug dose is,the more obvious the effect is.
引文
[1]沈娟,解丽培,汪文杰,等. ROC曲线评价TCT联合SCC-Ag水平测定诊断宫颈癌的价值[J].上海预防医学,2015,27(4):189-191.
[2]魏丽惠,赵超.宫颈癌及其癌前病变的筛查研究进展[J].中华妇幼临床医学杂志(电子版),2016,12(1):16-19.
[3]邓晰月,洪文旭,吴运妙,等.姜黄素对He La细胞增殖、迁移及miRNA表达谱的影响[J].中国妇幼保健,2015,30(26):4554-4558.
[4]李燕,张健.细胞与分子生物学常用实验技术[M].西安:第四军医大学出版社,2009:8-11,62-75.
[5]高岿然,史铁梅,邸晶,等.宫颈癌移植瘤大鼠动物模型的建立[J].中国组织工程研究,2008,12(28):5494-5498.
[6]秦川.实验动物学[M].北京:人民卫生出版社,2010:274-278.
[7]国家食品药品监督管理局.细胞毒类抗肿瘤药物非临床研究技术指导原则[J].中国新药与临床杂志,2008,27(6):462-465.
[8]王兰兰,许化溪.临床免疫学检验[M].北京:人民卫生出版社,2012:103-108,145-147.
[9]贺帅,吴彦萍,蔡湘仪,等.姜黄素对宫颈癌细胞生长与增殖的影响研究[J].现代生物医学进展,2017,17(1):7-10.
[10]田晓琳,杨臻,王建英,等.微小RNA与肿瘤的关系[J].癌症进展,2016,14(1):22-25.
[11]杨朝晖,章辉,顾华敏,等.宫颈癌组织中miR-21和Stat3的表达及相关性研究[J].浙江医学,2017,39(2):93-96.
[2]魏丽惠,赵超.宫颈癌及其癌前病变的筛查研究进展[J].中华妇幼临床医学杂志(电子版),2016,12(1):16-19.
[3]邓晰月,洪文旭,吴运妙,等.姜黄素对He La细胞增殖、迁移及miRNA表达谱的影响[J].中国妇幼保健,2015,30(26):4554-4558.
[4]李燕,张健.细胞与分子生物学常用实验技术[M].西安:第四军医大学出版社,2009:8-11,62-75.
[5]高岿然,史铁梅,邸晶,等.宫颈癌移植瘤大鼠动物模型的建立[J].中国组织工程研究,2008,12(28):5494-5498.
[6]秦川.实验动物学[M].北京:人民卫生出版社,2010:274-278.
[7]国家食品药品监督管理局.细胞毒类抗肿瘤药物非临床研究技术指导原则[J].中国新药与临床杂志,2008,27(6):462-465.
[8]王兰兰,许化溪.临床免疫学检验[M].北京:人民卫生出版社,2012:103-108,145-147.
[9]贺帅,吴彦萍,蔡湘仪,等.姜黄素对宫颈癌细胞生长与增殖的影响研究[J].现代生物医学进展,2017,17(1):7-10.
[10]田晓琳,杨臻,王建英,等.微小RNA与肿瘤的关系[J].癌症进展,2016,14(1):22-25.
[11]杨朝晖,章辉,顾华敏,等.宫颈癌组织中miR-21和Stat3的表达及相关性研究[J].浙江医学,2017,39(2):93-96.