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PKC iota promotes cellular proliferation by accelerated G1/S transition via interaction with CDK7 in esophageal squamous cell carcinoma

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Tumor Biology

Abstract

Protein kinase C iota (PKCι) has been shown to play an important role in tumorigenesis of many cancers. It was reported that frequent amplification and overexpression of PKCi were correlated with resistance to anoikis in primary esophageal squamous cell carcinomas (ESCC). In this study, we clarified a novel role of PKCι on the cell cycle progression and proliferation in ESCC. Western blot and immunohistochemistry (IHC) analysis showed that the expression of PKCι was higher in ESCC tumor tissues and cell lines. Meanwhile, IHC stain revealed that PKCι was positively correlated with clinical pathologic variables such as tumor size, tumor grade, and tumor invasion, as well as ki67. Immunoprecipitation and immunofluorescence assay revealed that PKCι/CDK7 has the physical interaction and were co-located in the cell nucleus. And this direct interaction could increase the phosphorylation level of CDK7. In vitro studies such as starvation and refeeding assay along with PKCι-shRNA transfection assay demonstrated that PKCι expression promoted proliferation of ESCC cells. And knocking PKCi down by silencing RNA (siRNA) significantly caused cell cycle arrest at G0/G1 phase, decreased rate of colony formation, and alleviated cellular apoptosis. This research provide new insights into PKCi signaling to more deeply understand its cancer-promoting function in ESCC.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (no. 81171140, no. 81472272).

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Authors and Affiliations

  1. Department of Medical Oncology, Affiliated Hospital of Nantong University, 20 Xisi Road, Nantong, Jiangsu, 226001, China

    Sujie Ni, Mei Li, Jialin Cheng, Li Liang & Wenyan Jiang

  2. Department of Gastroenterology, Affiliated Hospital of Nantong University, 20 Xisi Road, Nantong, Jiangsu, 226001, China

    Lingling Chen, Weijuan Zhao, Xiaohang Shan, Miaomiao Wu, Yayun Wang & Runzhou Ni

  3. Department of Pathology, Affiliated Hospital of Nantong University, 20 Xisi Road, Nantong, Jiangsu, 226001, China

    Jianguo Zhang

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  1. Sujie Ni
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  2. Lingling Chen
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  3. Mei Li
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  4. Weijuan Zhao
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  5. Xiaohang Shan
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  10. Wenyan Jiang
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  11. Jianguo Zhang
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  12. Runzhou Ni
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Corresponding authors

Correspondence to Jianguo Zhang or Runzhou Ni.

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Sujie Ni and Lingling Chen contributed equally to this work.

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Fig. S1

Silencing of PKCι in ECA 109 cells. ECA 109 cells were transfected with shRNA targeting either four PKCι or a scrambled sequence (control shRNA) for 48 h. Western blot analysis was performed to evaluate the expression of PKCι. The data showed the PKCι protein levels evidently reduced both in ECA109 cells following PKCι-shRNA#4 when compared with control shRNA transfected cells. (JPEG 4 kb)

High resolution image (TIFF 444 kb)

Fig. S2

Immunofluorescence assay was again performed to determine the Cellular localization of PKCι with CDK7 after expression of PKCι was resuced. Eca109 cells were transfected with PKCι-shRNA#4 for 48 h. Then, Immunofluorescence assay was again performed The result showed that the expression of aPKCi was obviously reduced, which especially in the nuclear, resulting in few co-localization within aPKCi and CDK7. (JPEG 15 kb)

High resolution image (TIFF 4.81 mb)

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Ni, S., Chen, L., Li, M. et al. PKC iota promotes cellular proliferation by accelerated G1/S transition via interaction with CDK7 in esophageal squamous cell carcinoma. Tumor Biol. 37, 13799–13809 (2016). https://doi.org/10.1007/s13277-016-5193-9

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  • DOI: https://doi.org/10.1007/s13277-016-5193-9

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