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Synaptonemal complex protein 3 is a prognostic marker in cervical cancer.

Cho H, Noh KH, Chung JY, Takikita M, Chung EJ, Kim BW, Hewitt SM, Kim TW, Kim JH - PLoS ONE (2014)

Bottom Line: Functional studies using NIH3T3 cells demonstrated that the C-terminal region of human SCP3 is important for AKT activation and its oncogenic potential.High expression of SCP3 was significantly associated with tumor stage (P = 0.002) and tumor grade (P<0.001), while SCP3 expression was positively associated with pAKT protein level in cervical neoplasias.Survival times for patients with cervical cancer overexpressing both SCP3 and pAKT (median, 134.0 months, n = 68) were significantly shorter than for patients with low expression of either SCP3 or pAKT (161.5 months, n = 108) as determined by multivariate analysis (P = 0.020).

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea; Institute of Women's Life Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea.

ABSTRACT
Synaptonemal complex protein 3 (SCP3), a member of Cor1 family, is up-regulated in various cancer cells; however, its oncogenic potential and clinical significance has not yet been characterized. In the present study, we investigated the oncogenic role of SCP3 and its relationship with phosphorylated AKT (pAKT) in cervical neoplasias. The functional role of SCP3 expression was investigated by overexpression or knockdown of SCP3 in murine cell line NIH3T3 and human cervical cancer cell lines CUMC6, SiHa, CaSki, and HeLa both in vitro and in vivo. Furthermore, we examined SCP3 expression in tumor specimens from 181 cervical cancer and 400 cervical intraepithelial neoplasia (CIN) patients by immunohistochemistry and analyzed the correlation between SCP3 expression and clinicopathologic factors or survival. Overexpression of SCP3 promoted AKT-mediated tumorigenesis both in vitro and in vivo. Functional studies using NIH3T3 cells demonstrated that the C-terminal region of human SCP3 is important for AKT activation and its oncogenic potential. High expression of SCP3 was significantly associated with tumor stage (P = 0.002) and tumor grade (P<0.001), while SCP3 expression was positively associated with pAKT protein level in cervical neoplasias. Survival times for patients with cervical cancer overexpressing both SCP3 and pAKT (median, 134.0 months, n = 68) were significantly shorter than for patients with low expression of either SCP3 or pAKT (161.5 months, n = 108) as determined by multivariate analysis (P = 0.020). Our findings suggest that SCP3 plays an important role in the progression of cervical cancer through the AKT signaling pathway, supporting the possibility that SCP3 may be a promising novel cancer target for cervical cancer therapy.

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hSCP3 increases oncogenic potentials of NIH3T3 cells in vitro and in vivo.(A) Western blot analysis of expression of hSCP3 in NIH3T3 cells retrovirally transduced with a pMSCV vector encoding hSCP3 (NIH3T3/hSCP3). NIH3T3/No insert cells were used as a control. Numbers below western blots refer to the relative values of the intensity normalized to no insert control. (B) In vitro growth curves of NIH3T3/hSCP3 cells. Cells were counted after trypan blue staining to exclude dead cells. (C) Colony-forming capacity of NIH3T3/hSPC3 cells in soft agar; (left) Representative images of average colony size in each group; (right) Bar graph representing the number of colonies with diameters greater than 2 mm in soft agar (scale bar: 1 mm). (D) Tumorigenicity of NIH3T3/hSCP3 cells. Balb/c Nude mice (n = 5) were inoculated subcutaneously with 1×105 cells/mouse of NIH3T3/no insert or NIH3T3/hSCP3 cells. Error bars represent the mean ± SD.
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pone-0098712-g001: hSCP3 increases oncogenic potentials of NIH3T3 cells in vitro and in vivo.(A) Western blot analysis of expression of hSCP3 in NIH3T3 cells retrovirally transduced with a pMSCV vector encoding hSCP3 (NIH3T3/hSCP3). NIH3T3/No insert cells were used as a control. Numbers below western blots refer to the relative values of the intensity normalized to no insert control. (B) In vitro growth curves of NIH3T3/hSCP3 cells. Cells were counted after trypan blue staining to exclude dead cells. (C) Colony-forming capacity of NIH3T3/hSPC3 cells in soft agar; (left) Representative images of average colony size in each group; (right) Bar graph representing the number of colonies with diameters greater than 2 mm in soft agar (scale bar: 1 mm). (D) Tumorigenicity of NIH3T3/hSCP3 cells. Balb/c Nude mice (n = 5) were inoculated subcutaneously with 1×105 cells/mouse of NIH3T3/no insert or NIH3T3/hSCP3 cells. Error bars represent the mean ± SD.

Mentions: A previous study reported that human SCP3 (hSCP3) is expressed in various cancer cells and that its expression in human specimens is associated with oncogenesis [8], [10]. However, the oncogenic potential of SCP3 has not been sufficiently characterized at either the molecular or cellular level. To examine whether hSCP3 may have cellular tumorigenic potential, we first used a non-tumorigenic murine fibroblast NIH3T3 cell line and established the cell lines expressing hSCP3 using a retroviral transduction system (NIH3T3/hSCP3). NIH3T3/no insert (empty vector) cells were also established as a control. The expression of SCP3 protein in NIH3T3/hSCP3 cells was confirmed by Western blot (Figure 1A). Cell proliferation of transduced NIH3T3 cells was measured by counting the number of live cells after trypan blue staining. In cell proliferation assays (Figure 1B), NIH3T3/hSCP3 cells exhibited a significantly increased proliferation rate compared with NIH3T3/no insert cells (P<0.005). The results of a soft agar colony formation assay with the same cell lines were consistent with those of the cell proliferation assay (Figure 1C). NIH3T3/hSCP3 cells exhibited significantly higher potentials of colony formation in soft agar as compared with NIH3T3/no insert cells (P<0.005). We next subcutaneously injected hSCP3- or no insert-expressing NIH3T3 cell in the left flank of athymic Balb/c Nude mice (Figure 1D). Consistent with our in vitro data, the tumor-forming ability of NIH3T3 cells was drastically increased by ectopically-expressing hSCP3 (no insert vs. hSCP3, P<0.001). We next explored whether other genes from the murine Cor1 family such mXLR, mXMR and mSCP3, which share more than 60% homology with hSCP3, could have similar tumorigenic potential in the NIH3T3 model [10]. As shown in Figure S2, ectopic expression of Cor1 family members substantially promoted in vitro cell proliferation and soft agar colony formation as well as in vivo tumor formation in NIH3T3 cell line model, which was consistent with that of the hSCP3. Taken together, these data clearly demonstrate the oncogenic potentials of SCP3 and other Cor1 members in NIH3T3 model.


Synaptonemal complex protein 3 is a prognostic marker in cervical cancer.

Cho H, Noh KH, Chung JY, Takikita M, Chung EJ, Kim BW, Hewitt SM, Kim TW, Kim JH - PLoS ONE (2014)

hSCP3 increases oncogenic potentials of NIH3T3 cells in vitro and in vivo.(A) Western blot analysis of expression of hSCP3 in NIH3T3 cells retrovirally transduced with a pMSCV vector encoding hSCP3 (NIH3T3/hSCP3). NIH3T3/No insert cells were used as a control. Numbers below western blots refer to the relative values of the intensity normalized to no insert control. (B) In vitro growth curves of NIH3T3/hSCP3 cells. Cells were counted after trypan blue staining to exclude dead cells. (C) Colony-forming capacity of NIH3T3/hSPC3 cells in soft agar; (left) Representative images of average colony size in each group; (right) Bar graph representing the number of colonies with diameters greater than 2 mm in soft agar (scale bar: 1 mm). (D) Tumorigenicity of NIH3T3/hSCP3 cells. Balb/c Nude mice (n = 5) were inoculated subcutaneously with 1×105 cells/mouse of NIH3T3/no insert or NIH3T3/hSCP3 cells. Error bars represent the mean ± SD.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4048308&req=5

pone-0098712-g001: hSCP3 increases oncogenic potentials of NIH3T3 cells in vitro and in vivo.(A) Western blot analysis of expression of hSCP3 in NIH3T3 cells retrovirally transduced with a pMSCV vector encoding hSCP3 (NIH3T3/hSCP3). NIH3T3/No insert cells were used as a control. Numbers below western blots refer to the relative values of the intensity normalized to no insert control. (B) In vitro growth curves of NIH3T3/hSCP3 cells. Cells were counted after trypan blue staining to exclude dead cells. (C) Colony-forming capacity of NIH3T3/hSPC3 cells in soft agar; (left) Representative images of average colony size in each group; (right) Bar graph representing the number of colonies with diameters greater than 2 mm in soft agar (scale bar: 1 mm). (D) Tumorigenicity of NIH3T3/hSCP3 cells. Balb/c Nude mice (n = 5) were inoculated subcutaneously with 1×105 cells/mouse of NIH3T3/no insert or NIH3T3/hSCP3 cells. Error bars represent the mean ± SD.
Mentions: A previous study reported that human SCP3 (hSCP3) is expressed in various cancer cells and that its expression in human specimens is associated with oncogenesis [8], [10]. However, the oncogenic potential of SCP3 has not been sufficiently characterized at either the molecular or cellular level. To examine whether hSCP3 may have cellular tumorigenic potential, we first used a non-tumorigenic murine fibroblast NIH3T3 cell line and established the cell lines expressing hSCP3 using a retroviral transduction system (NIH3T3/hSCP3). NIH3T3/no insert (empty vector) cells were also established as a control. The expression of SCP3 protein in NIH3T3/hSCP3 cells was confirmed by Western blot (Figure 1A). Cell proliferation of transduced NIH3T3 cells was measured by counting the number of live cells after trypan blue staining. In cell proliferation assays (Figure 1B), NIH3T3/hSCP3 cells exhibited a significantly increased proliferation rate compared with NIH3T3/no insert cells (P<0.005). The results of a soft agar colony formation assay with the same cell lines were consistent with those of the cell proliferation assay (Figure 1C). NIH3T3/hSCP3 cells exhibited significantly higher potentials of colony formation in soft agar as compared with NIH3T3/no insert cells (P<0.005). We next subcutaneously injected hSCP3- or no insert-expressing NIH3T3 cell in the left flank of athymic Balb/c Nude mice (Figure 1D). Consistent with our in vitro data, the tumor-forming ability of NIH3T3 cells was drastically increased by ectopically-expressing hSCP3 (no insert vs. hSCP3, P<0.001). We next explored whether other genes from the murine Cor1 family such mXLR, mXMR and mSCP3, which share more than 60% homology with hSCP3, could have similar tumorigenic potential in the NIH3T3 model [10]. As shown in Figure S2, ectopic expression of Cor1 family members substantially promoted in vitro cell proliferation and soft agar colony formation as well as in vivo tumor formation in NIH3T3 cell line model, which was consistent with that of the hSCP3. Taken together, these data clearly demonstrate the oncogenic potentials of SCP3 and other Cor1 members in NIH3T3 model.

Bottom Line: Functional studies using NIH3T3 cells demonstrated that the C-terminal region of human SCP3 is important for AKT activation and its oncogenic potential.High expression of SCP3 was significantly associated with tumor stage (P = 0.002) and tumor grade (P<0.001), while SCP3 expression was positively associated with pAKT protein level in cervical neoplasias.Survival times for patients with cervical cancer overexpressing both SCP3 and pAKT (median, 134.0 months, n = 68) were significantly shorter than for patients with low expression of either SCP3 or pAKT (161.5 months, n = 108) as determined by multivariate analysis (P = 0.020).

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea; Institute of Women's Life Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea.

ABSTRACT
Synaptonemal complex protein 3 (SCP3), a member of Cor1 family, is up-regulated in various cancer cells; however, its oncogenic potential and clinical significance has not yet been characterized. In the present study, we investigated the oncogenic role of SCP3 and its relationship with phosphorylated AKT (pAKT) in cervical neoplasias. The functional role of SCP3 expression was investigated by overexpression or knockdown of SCP3 in murine cell line NIH3T3 and human cervical cancer cell lines CUMC6, SiHa, CaSki, and HeLa both in vitro and in vivo. Furthermore, we examined SCP3 expression in tumor specimens from 181 cervical cancer and 400 cervical intraepithelial neoplasia (CIN) patients by immunohistochemistry and analyzed the correlation between SCP3 expression and clinicopathologic factors or survival. Overexpression of SCP3 promoted AKT-mediated tumorigenesis both in vitro and in vivo. Functional studies using NIH3T3 cells demonstrated that the C-terminal region of human SCP3 is important for AKT activation and its oncogenic potential. High expression of SCP3 was significantly associated with tumor stage (P = 0.002) and tumor grade (P<0.001), while SCP3 expression was positively associated with pAKT protein level in cervical neoplasias. Survival times for patients with cervical cancer overexpressing both SCP3 and pAKT (median, 134.0 months, n = 68) were significantly shorter than for patients with low expression of either SCP3 or pAKT (161.5 months, n = 108) as determined by multivariate analysis (P = 0.020). Our findings suggest that SCP3 plays an important role in the progression of cervical cancer through the AKT signaling pathway, supporting the possibility that SCP3 may be a promising novel cancer target for cervical cancer therapy.

Show MeSH
Related in: MedlinePlus