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Src family kinase inhibitor PP2 efficiently inhibits cervical cancer cell proliferation through down-regulating phospho-Src-Y416 and phospho-EGFR-Y1173

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Abstract

Tyrosine (Y) kinases inhibitors have been approved for targeted treatment of cancer. However, their clinical use is limited to some cancers and the mechanism of their action remains unclear. Previous study has indicated that PP2, a selective inhibitor of the Src family of non-receptor tyrosine kinases (nRTK), efficiently repressed cervical cancer growth in vitro and in vivo. In this regard, our aims are to explore the mechanism of PP2 on cervical cancer cell growth inhibition by investigating the suppressive divergence among PP1, PP2, and a negative control compound PP3. MTT results showed that three compounds had different inhibitory effects on proliferation of two cervical cancer cells, HeLa and SiHa, and PP2 was most efficient in a time- and dose-dependent manner. Moreover, we found 10 μM PP2 down-regulated pSrc-Y416 (P < 0.05), pEGFR-Y845 (P < 0.05), and -Y1173 (P < 0.05) expression levels, while 10 μM PP1 down-regulated pSrc-Y416 (P < 0.05) and pEGFR-Y845 (P < 0.05), but not pEGFR-Y1173; 10 μM PP3 down-regulated only pEGFR-Y1173 (P < 0.05). PP2 could modulate cell cycle arrest by up-regulating p21Cip1 and p27Kip1 in both HeLa and SiHa cells and down-regulating expression of cyclin A, and cyclin dependent kinase-2, -4 (Cdk-2, -4) in HeLa and of cyclin B and Cdk-2 in SiHa. Our results indicate that Src pathway and EGFR pathway play different roles in the proliferation of cervical cancer cells and PP2 efficiently reduces cervical cancer cell proliferation by reduction of both Src and EGFR activity.

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Abbreviations

nrTKs:

Non-receptor tyrosine kinases

Y:

Tyrosine

Src:

Sarcoma

EGFR:

Epidermal growth factor receptor

PP1:

4-Amino-5-(4-methylphenyl)-7-(t-butyl) pyrazolo[3,4-d]pyrimidine

PP2:

4-Amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo[3,4-d] pyrimidine

PP3:

4-Amino-7-phenylpyrazol[3,4-d] pyrimidine

Cdk:

Cyclin-dependent kinase

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

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Acknowledgments

This work was supported by Major State Basic Research Development Programs (2009CB521800; 2010CB529400) and the National Natural Sciences Foundation of China (30672422). We thank Ms. Xingcui Wang for the flow cytometry work.

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None.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Cancer Institute, Capital Medical University, No.10 Xitoutiao, You An Men, Beijing, 100069, China

    Lu Kong, Zhihong Deng, Haiying Shen & Yuxiang Zhang

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  1. Lu Kong
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  2. Zhihong Deng
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  3. Haiying Shen
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  4. Yuxiang Zhang
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Corresponding author

Correspondence to Yuxiang Zhang.

Additional information

Lu Kong and Zhihong Deng contributed equally to this study.

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Kong, L., Deng, Z., Shen, H. et al. Src family kinase inhibitor PP2 efficiently inhibits cervical cancer cell proliferation through down-regulating phospho-Src-Y416 and phospho-EGFR-Y1173. Mol Cell Biochem 348, 11–19 (2011). https://doi.org/10.1007/s11010-010-0632-1

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