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犬乳腺肿瘤是犬类的一种常见肿瘤性疾病,目前治疗多为手术结合放、化疗法,但结果多易转移复发。双氢青蒿素(dihydroartemisinin,DHA)是中药提取物,近年来其抗肿瘤作用越来越受到重视。为探求双氢青蒿素是否影响犬乳腺肿瘤细胞的侵袭及其影响机制,本试验取对数增长期状态相同的犬乳腺肿瘤细胞CHMm,使用低(5μM)、中(10μM)、高(20μM)浓度DHA处理并分别培养24,48,72h。收集各组细胞,使用CCK-8法检测细胞活性;利用细胞划痕试验检测细胞迁移性;利用transwell检测细胞侵袭性;利用CCK-8检测细胞毒性。结果表明:DHA对CHMm细胞抑制率呈明显药物浓度依赖性。细胞划痕试验检测细胞迁移速率结果表明,相同培养时间,与空白对照组相比,低、中、高试验组对CHMm细胞迁移性抑制作用效果极显著(p<0.01)。各组组间比较,DHA对细胞迁移性抑制影响差异极显著(p<0.01)。DHA对CHMm细胞迁移性抑制呈明显的药物时间、浓度依赖性。细胞侵袭试验检测细胞侵袭性结果表明,相同培养时间,与空白对照组相比,低、中、高试验组穿透基底膜着色细胞个数显著增多,说明DHA对CHMm细胞侵袭性抑制效果极显著(p<0.01),组间比较,不同DHA作用浓度效果差异显著(p<0.05)。相同DHA作用浓度,随着培养时间的增加,细胞侵袭性降低,效果显著(p<0.05)。细胞毒性结果表明,相同培养时间,与空白对照组相比,低、中、高试验组对CHMm细胞抑制性影响效果极显著(p<0.01)。各组组间比较,DHA对细胞抑制性影响差异极显著(p<0.01)。综上所述,DHA对犬乳腺肿瘤细胞增殖、迁移和侵袭作用具有显著抑制作用,且随着DHA作用浓度增加和时间增长,抑制作用增强,呈浓度时间依赖性。
Canine mammary tumors are a common tumor of dogs, the current treatment of surgery combined with radiotherapy and chemotherapy, but the results are more likely to relapse. Dihydroartemisinin (DHA) is a traditional Chinese medicine extract, and its antitumor effect has drawn more and more attention in recent years. In order to find out whether dihydroartemisinin affects the invasion of mammary tumor cells and its mechanism, we use low (5μM), moderate (10μM), high (20μM ) Concentration of DHA treatment and were cultured for 24,48,72h. The cells in each group were collected and the cell viability was determined by CCK-8 assay. Cell migration was assayed by cell scratch assay. Cell invasion was assayed by transwell assay. Cytotoxicity was detected by CCK-8 assay. The results showed that: DHA inhibition rate of CHMm cells was significantly drug concentration-dependent. The results of cell scratch assay showed that the effect of inhibiting the migration of CHMm cells was significant (p <0.01) in the low, middle and high test groups compared with the blank control group at the same culture time. The differences between groups were significant (P <0.01). DHA on the migration inhibition of CHMm cells showed a significant drug time and concentration-dependent. Cell invasion test to detect cell invasive results showed that the same culture time, compared with the blank control group, low, medium and high test group significantly increased the number of penetrating basement membrane staining cells, indicating that DHA CHMm cell invasive inhibitory effect was significant (p <0.01). There was significant difference in effect of different concentration of DHA between two groups (p <0.05). The same concentration of DHA, with the increase of culture time, cell invasion decreased, the effect was significant (p <0.05). Cytotoxicity results showed that the inhibitory effect of low, middle and high test groups on CHMm cells was significant (p <0.01) at the same incubation time compared with the blank control group. The differences of cytostatic effects between DHA groups were significant (p <0.01). In summary, DHA had a significant inhibitory effect on the proliferation, migration and invasion of mammary tumor cells. With the increase of DHA concentration and time, the inhibitory effect was enhanced in a dose-and-time-dependent manner.