Danshen Injection Inhibits Platelets-induced Metastasis<italic style="font-style: italic"> </italic>of Breast Cancer Cells<italic style="font-style: italic"> In Vitro</italic>

TIAN Huiru; JIANG Siqin; LYU Hong; ZHUO Dongliang; FU Weiran; FU Jianjiang

doi:10.13422/j.cnki.syfjx.20230325

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Danshen Injection Inhibits Platelets-induced Metastasis of Breast Cancer Cells In Vitro

更新时间：2023-10-13

- Danshen Injection Inhibits Platelets-induced Metastasis of Breast Cancer Cells In Vitro
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 21, Pages: 79-85(2023)
- 作者机构：
  
  1.江西中医药大学，南昌 330004
  2.首都医科大学基础医学院，北京 100069
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230325
  CLC： R22;R242;R2-031;R285.5
- Published：05 November 2023，
  
  Published Online：14 December 2022，
  
  Received：05 September 2022，
- 稿件说明：
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田会茹,姜思琴,吕红等.丹参注射液抑制血小板诱导的乳腺癌细胞体外转移作用[J].中国实验方剂学杂志,2023,29(21):79-85.

TIAN Huiru,JIANG Siqin,LYU Hong,et al.Danshen Injection Inhibits Platelets-induced Metastasisof Breast Cancer Cells In Vitro[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(21):79-85.
田会茹,姜思琴,吕红等.丹参注射液抑制血小板诱导的乳腺癌细胞体外转移作用[J].中国实验方剂学杂志,2023,29(21):79-85. DOI： 10.13422/j.cnki.syfjx.20230325.

TIAN Huiru,JIANG Siqin,LYU Hong,et al.Danshen Injection Inhibits Platelets-induced Metastasisof Breast Cancer Cells In Vitro[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(21):79-85. DOI： 10.13422/j.cnki.syfjx.20230325.

摘要

目的

观察丹参注射液对血小板（PLT）诱导的乳腺癌细胞体外转移作用。

方法

采用噻唑蓝（MTT）比色法观察丹参注射液对MDA-MB-231细胞体外生长的影响；采用Oris

体外迁移试剂盒检测丹参注射液（终质量浓度分别为4、8、16 g·L

-1

）对PLT（1.5×10

个/L）诱导的乳腺癌细胞体外迁移的影响；采用Transwell小室法检测丹参注射液对PLT诱导的细胞侵袭的影响；采用细胞免疫荧光法和蛋白免疫印迹法（Western blot）检测丹参注射液对PLT诱导的上皮间质转化（EMT）关键性转录因子Slug、Snail蛋白表达的影响；采用酶联免疫吸附测定法（ELISA）检测丹参注射液（终质量浓度为4、8、16、32、64 g·L

-1

）对转化生长因子-

（TGF-

）分泌的影响；采用Western blot观察丹参注射液对PLT诱导MDA-MB-231细胞Podoplain（PDPN）蛋白表达的影响。

结果

与空白组比较，丹参注射液组（32、64 g·L

-1

）细胞的吸光度

570

均明显降低（

0.05，

0.01）；丹参注射液组（4、8、16 g·L

-1

）

570

差异无统计学意义。与空白组比较，PLT组迁移、侵袭细胞数明显增加，丹参注射液各组则可明显抑制PLT诱导的细胞迁移和侵袭。与空白组比较，PLT组细胞E-钙黏蛋白（E-cadherin）表达明显降低，丹参注射液可明显逆转PLT的这一作用。与空白组比较，PLT组Slug、Snail蛋白表达明显升高（

0.05，

0.01），丹参注射液则明显逆转PLT诱导的Snail蛋白表达（

0.05，

0.01）；PLT组TGF-

含量显著升高（

0.01），丹参注射液组（16、32、64 g·L

-1

）可明显降低PLT诱导的TGF-

分泌（

0.05，

0.01），其余丹参注射液组对TGF-

分泌无明显影响；PLT组细胞PDPN蛋白表达显著升高（

0.01），丹参注射液可显著抑制PLT诱导的PDPN表达升高（

0.01）。

结论

丹参注射液可抑制PLT诱导的乳腺癌细胞的迁移、侵袭和EMT作用，其机制可能是通过下调乳腺癌细胞PDPN表达、干扰PLT与肿瘤细胞之间直接作用与PLT分泌TGF-

作用无关。

Abstract

Objective

To observe the effect of Danshen injection （DAN） on platelet （PLT）-induced metastasis of breast cancer cells

in vitro

Method

The 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide （MTT） assay was used to observe the effect of DAN on the growth of MDA-MB-231 cells

in vitro

. Oris™ migration assay was used to determine the effect of DAN （final mass concentrations 4， 8， 16 g·L

-1

） on PLT （1.5×10

cells/L）-induced migration of breast cancer cells

in vitro

. The effect of DAN on PLT-induced cell invasion was detected by Transwell assay. Immunofluorescence and Western blot were used to detect the effect of DAN on the protein expression associated with PLT-induced epithelial-mesenchymal transition （EMT）. In addition， enzyme-linked immune-sorbent assay （ELISA） was used to determine the effect of DAN （final mass concentrations 4， 8， 16， 32， 64 g·L

-1

） on the secretion of transforming growth factor-

（TGF-

）. Western blot was used to observe the effect of DAN on the expression of podoplanin （PDPN） protein in MDA-MB-231 cells induced by PLT.

Result

Compared with the blank group， the DAN groups （32 and 64 g·L

-1

） showed decreased

570

（

0.05，

0.01）， and there was no significant difference in

570

between DAN groups （4， 8， 16 g·L

-1

）. Compared with the blank group， the PLT group showed increased cell migration and invasion， while DAN groups significantly inhibited PLT-induced cell migration and invasion. Compared with the blank group， the PLT group showed decreased expression of E-cadherin， while DAN could significantly reverse this effect of PLT. Compared with the blank group， the PLT group showed increased Slug and Snail protein expression （

0.05，

0.01）， while DAN significantly reversed Snail protein expression induced by PLT （

0.05，

0.01）. The content of TGF-

in the PLT group increased （

0.01）， while the secretion of TGF-

induced by PLT decreased in the DAN groups （16， 32， and 64 g·L

-1

）（

0.05，

0.01）， and the secretion of TGF-

was not significantly affected in other DAN groups. PDPN protein expression in the PLT group increased （

0.01）， while DAN could significantly inhibit PLT-induced PDPN expression （

0.01）.

Conclusion

DAN can inhibit PLT-induced migration， invasion， and EMT of breast cancer cells. The mechanism may be related to the direct action between breast cancer cells and tumor cells by down-regulating PDPN expression and interfering with PLT and has nothing to do with the effect of TGF-

secretion of PLT.

关键词

丹参注射液血小板乳腺癌转移

Keywords

Danshen injectionplateletbreast cancermetastasis

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Danshen Injection Inhibits Platelets-induced Metastasis of Breast Cancer Cells In Vitro

DOI：10.13422/j.cnki.syfjx.20230325

摘要

Abstract

关键词

Keywords

references