灵芝酸A对乳腺癌MCF-7细胞迁移和对激酶插入区受体基因表达的影响
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摘要
目的研究灵芝酸A对乳腺癌MCF-7细胞迁移及激酶插入区受体(KDR)基因表达的影响。方法将对数生长期乳腺癌MCF-7细胞分为空白组、对照组和低、高2个剂量实验组。空白组用改良杜氏伊格尔培养基100μL处置;对照组用0. 25 mg·L~(-1)的表柔比星溶液100μL处置;低、高2个剂量实验组用0. 1,0. 5 mmol·L~(-1)的灵芝酸A溶液100μL处置。用噻唑蓝比色法检测各组乳腺癌MCF-7细胞增殖抑制率,用细胞划痕实验检测乳腺癌MCF-7细胞迁移能力,用逆转录-聚合酶链反应检测各组乳腺癌MCF-7细胞KDR mRNA表达水平,用免疫印迹法检测各组乳腺癌MCF-7细胞KDR蛋白表达水平。结果处置24 h后,空白组、低剂量实验组、高剂量实验组和对照组的MCF-7细胞增殖抑制率分别是(0. 00)%,(39. 57±5. 29)%,(71. 37±7. 35)%和(72. 03±7. 64)%;这4组的细胞愈合率分别是(73. 27±8. 66)%,(52. 48±6. 54)%,(18. 73±2. 31)%和(18. 35±2. 48)%;这4组的KDR mRNA相对表达量分别是0. 95±0. 12,0. 65±0. 07,0. 21±0. 02和0. 20±0. 01;这4组的KDR蛋白相对表达量分别是1. 05±0. 13,0. 81±0. 10,0. 43±0. 05和0. 41±0. 05,3个给药组与空白组比较或高剂量实验组和对照组与低剂量实验组比较,差异均有统计学意义(均P <0. 05)。结论灵芝酸A能剂量依赖性地抑制乳腺癌MCF-7细胞增殖,有效降低细胞迁移能力,其作用可能与下调KDR mRNA及KDR蛋白表达有关。
Objective To analyze the effects of ganoderic acid A on cell migration and expression of kinase insert domain containing receptor( KDR) gene in breast cancer MCF-7 cells. Methods Breast cancer MCF-7 cells in logarithmic growth phase were divided into 4 groups:blank group,experimental-L group,experimental-H group and control group. The blank group was treated with dulbecco's modified eagle medium 100 L; the experimental-L,experimental-H groups were treated with 0. 1,0. 5 mmol·L~(-1) ganoderic acid A solution 100 μL,the control group was treated with 0. 25 mmol·L~(-1) epirubicin solution 100 μL. The breast cancer MCF-7 cell proliferation inhibition rate was detected by methylthiazoltetrazolium colorimetry. The cell wound scratch assay was used to detect the migration ability of breast cancer MCF-7 cells. The expression level of KDR mRNA were detected by reverse transcription-polymerase chain reaction. The expression levels of KDR protein were detected by Western blot. Results After 24 h of disposal,the healing rates of breast cancer MCF-7 cells in blank group,experimental-L group,experimental-H group and control group were respectively( 0. 00) %,( 39. 57 ± 5. 29) %,( 71. 37 ± 7. 35) % and( 72. 03 ± 7. 64) %;the healing rates of cells in the 4 groups were respectively( 73. 27 ± 8. 66) %,( 52. 48 ± 6. 54) %,( 18. 73 ± 2. 31) %and( 18. 35 ± 2. 48) %; the relative expression of KDR mRNA in the 4 groups were respectively 0. 95 ± 0. 12,0. 65 ± 0. 07,0. 21 ± 0. 02 and 0. 20 ± 0. 01; the relative expression of KDR protein in the 4 groups were respectively1. 05 ± 0. 13,0. 81 ± 0. 10,0. 43 ± 0. 05 and 0. 41 ± 0. 05,comparing between three drugs groups with blank group,or comparison between experimental-H group and control group with experimental-L group,the differences were significant( all P < 0. 05). Conclusion Ganoderma lucidum acid A can inhibit the proliferation of breast cancer MCF-7 cells in a dose-dependent manner,effectively reduce cell migration ability,which may be related to the down-regulation of KDR mRNA and KDR protein expression.
Objective To analyze the effects of ganoderic acid A on cell migration and expression of kinase insert domain containing receptor( KDR) gene in breast cancer MCF-7 cells. Methods Breast cancer MCF-7 cells in logarithmic growth phase were divided into 4 groups:blank group,experimental-L group,experimental-H group and control group. The blank group was treated with dulbecco's modified eagle medium 100 L; the experimental-L,experimental-H groups were treated with 0. 1,0. 5 mmol·L~(-1) ganoderic acid A solution 100 μL,the control group was treated with 0. 25 mmol·L~(-1) epirubicin solution 100 μL. The breast cancer MCF-7 cell proliferation inhibition rate was detected by methylthiazoltetrazolium colorimetry. The cell wound scratch assay was used to detect the migration ability of breast cancer MCF-7 cells. The expression level of KDR mRNA were detected by reverse transcription-polymerase chain reaction. The expression levels of KDR protein were detected by Western blot. Results After 24 h of disposal,the healing rates of breast cancer MCF-7 cells in blank group,experimental-L group,experimental-H group and control group were respectively( 0. 00) %,( 39. 57 ± 5. 29) %,( 71. 37 ± 7. 35) % and( 72. 03 ± 7. 64) %;the healing rates of cells in the 4 groups were respectively( 73. 27 ± 8. 66) %,( 52. 48 ± 6. 54) %,( 18. 73 ± 2. 31) %and( 18. 35 ± 2. 48) %; the relative expression of KDR mRNA in the 4 groups were respectively 0. 95 ± 0. 12,0. 65 ± 0. 07,0. 21 ± 0. 02 and 0. 20 ± 0. 01; the relative expression of KDR protein in the 4 groups were respectively1. 05 ± 0. 13,0. 81 ± 0. 10,0. 43 ± 0. 05 and 0. 41 ± 0. 05,comparing between three drugs groups with blank group,or comparison between experimental-H group and control group with experimental-L group,the differences were significant( all P < 0. 05). Conclusion Ganoderma lucidum acid A can inhibit the proliferation of breast cancer MCF-7 cells in a dose-dependent manner,effectively reduce cell migration ability,which may be related to the down-regulation of KDR mRNA and KDR protein expression.
引文
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[10]于治灏,余岳,杨欣,等.沉默KDR基因乳腺癌MCF-7细胞的生物学特征改变[J].中国组织工程研究,2015,19(28):4514-4519.
[2]王颀,连臻强.中国乳腺癌筛查现状和评价[J].中华乳腺病杂志(电子版),2015,9(3):159-162.
[3]查良平,袁媛,黄璐琦.灵芝古今临床效用考[J].中国现代中药,2016,18(5):653-656.
[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.
[5]周蓉蓉,高鹏飞,张文意,等.槐耳板蓝根双向发酵提取物抑制乳腺癌MCF-7细胞迁移/侵袭及相关因子的研究[J].中国中医药信息杂志,2016,23(5):64-68.
[6]药立波.医学分子生物学实验技术[M].北京:人民卫生出版社,2014:303-305.
[7]李国华,李晔,梅锡玲,等.灵芝三萜类化合物研究进展[J].中草药,2015,46(12):1858-1862.
[8]刘海鹏,郑克彬,单小松.灵芝酸A对人胶质瘤细胞U251细胞增殖、凋亡和侵袭的影响[J].中国比较医学杂志,2016,26(3):64-69.
[9]潘恩山,李煜罡.灵芝酸A对人前列腺癌细胞DU-145细胞凋亡的影响[J].广东医学,2017,38(1):87-90.
[10]于治灏,余岳,杨欣,等.沉默KDR基因乳腺癌MCF-7细胞的生物学特征改变[J].中国组织工程研究,2015,19(28):4514-4519.