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Apoptosis inhibitor-5 overexpression is associated with tumor progression and poor prognosis in patients with cervical cancer.

Cho H, Chung JY, Song KH, Noh KH, Kim BW, Chung EJ, Ylaya K, Kim JH, Kim TW, Hewitt SM, Kim JH - BMC Cancer (2014)

Bottom Line: Immunohistochemical staining showed that API5 expression increased during the normal to tumor transition of cervical carcinoma (P < 0.001), and this increased expression was significantly associated with tumor stage (P = 0.004), tumor grade (P < 0.001), and chemo-radiation response (P = 0.004).In multivariate analysis, API5+ (P = 0.039) and combined API5+/pERK1/2+ (P = 0.032) were independent prognostic factors for overall survival.API5 expression is associated with pERK1/2 in a subset of cervical cancer patients and its expression predicts poor overall survival, supporting that API5 may be a promising novel target for therapeutic interventions.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Gangnam Severance Hospital, Yonsei University College of Medicine, 146-92 Dogok-Dong, Gangnam-Gu, Seoul 135-720, South Korea. genejock@helix.nih.gov.

ABSTRACT

Background: The apoptosis inhibitor-5 (API5), anti-apoptosis protein, is considered a key molecule in the tumor progression and malignant phenotype of tumor cells. Here, we investigated API5 expression in cervical cancer, its clinical significance, and its relationship with phosphorylated extracellular signal-regulated kinase 1 and 2 (pERK1/2) in development and progression of cervical cancer.

Methods: API5 effects on cell growth were assessed in cervical cancer cell lines. API5 and pERK1/2 immunohistochemical staining were performed on a cervical cancer tissue microarray consisting of 173 primary cervical cancers, 306 cervical intraepithelial neoplasias (CINs), and 429 matched normal tissues.

Results: API5 overexpression promoted cell proliferation and colony formation in CaSki cells, whereas API5 knockdown inhibited the both properties in HeLa cells. Immunohistochemical staining showed that API5 expression increased during the normal to tumor transition of cervical carcinoma (P < 0.001), and this increased expression was significantly associated with tumor stage (P = 0.004), tumor grade (P < 0.001), and chemo-radiation response (P = 0.004). API5 expression levels were positively associated with pERK1/2 in cervical cancer (P < 0.001) and high grade CIN (P = 0.031). In multivariate analysis, API5+ (P = 0.039) and combined API5+/pERK1/2+ (P = 0.032) were independent prognostic factors for overall survival.

Conclusions: API5 expression is associated with pERK1/2 in a subset of cervical cancer patients and its expression predicts poor overall survival, supporting that API5 may be a promising novel target for therapeutic interventions.

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Related in: MedlinePlus

API5 role in cell proliferation and colony formation of HEK293, CaSki, and HeLa cell lines. (A) API5 protein expression was analyzed by western blot. (B) Proliferation assay: 2 × 104 cells were plated in 24 well plates and cultured for additional 6 days. Cells were collected by trypsinization at the indicated times, and live cells were counted after trypan blue staining under a haemocytometer. (C) Colony formation: 500 cells were plated in 6 well plates and cultured for 2 weeks and formed colonies were stained with crystal violet. Data depicted as mean + s.e.m. from one representative experiment performed in triplicate.
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Fig2: API5 role in cell proliferation and colony formation of HEK293, CaSki, and HeLa cell lines. (A) API5 protein expression was analyzed by western blot. (B) Proliferation assay: 2 × 104 cells were plated in 24 well plates and cultured for additional 6 days. Cells were collected by trypsinization at the indicated times, and live cells were counted after trypan blue staining under a haemocytometer. (C) Colony formation: 500 cells were plated in 6 well plates and cultured for 2 weeks and formed colonies were stained with crystal violet. Data depicted as mean + s.e.m. from one representative experiment performed in triplicate.

Mentions: To evaluate the effects of API5 on cell proliferation, the API5 expression vector or the control vector (no insert) were transfected into CaSki cells which have a low level of API5 expression. Non-tumorigenic HEK 293 cells with low background of API5 expression level were used as a positive control for comparison. Conversely, siRNA targeting API5 (siAPI5) or GFP (irrelevant negative control, siGFP) were also transfected into HeLa cells which have a high level of API5 expression. API5 expression level in the transfected cells were detected by western blotting (Figure 2A). The cell growth assay revealed that cell growth rate in both of the API5-transfected cells was significantly higher than control groups (Figure 2B). Similar increase was also observed in colony formation assay (Figure 2C). In contrast, knock-down of API5 in HeLa cells significantly decreased both of the cell growth rate and colony formation efficacy compared with siGFP control group (Figure 2B and C). These data demonstrate that API5 has a key role in cell proliferation and colony formation of cervical cancer cells.Figure 2


Apoptosis inhibitor-5 overexpression is associated with tumor progression and poor prognosis in patients with cervical cancer.

Cho H, Chung JY, Song KH, Noh KH, Kim BW, Chung EJ, Ylaya K, Kim JH, Kim TW, Hewitt SM, Kim JH - BMC Cancer (2014)

API5 role in cell proliferation and colony formation of HEK293, CaSki, and HeLa cell lines. (A) API5 protein expression was analyzed by western blot. (B) Proliferation assay: 2 × 104 cells were plated in 24 well plates and cultured for additional 6 days. Cells were collected by trypsinization at the indicated times, and live cells were counted after trypan blue staining under a haemocytometer. (C) Colony formation: 500 cells were plated in 6 well plates and cultured for 2 weeks and formed colonies were stained with crystal violet. Data depicted as mean + s.e.m. from one representative experiment performed in triplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4125689&req=5

Fig2: API5 role in cell proliferation and colony formation of HEK293, CaSki, and HeLa cell lines. (A) API5 protein expression was analyzed by western blot. (B) Proliferation assay: 2 × 104 cells were plated in 24 well plates and cultured for additional 6 days. Cells were collected by trypsinization at the indicated times, and live cells were counted after trypan blue staining under a haemocytometer. (C) Colony formation: 500 cells were plated in 6 well plates and cultured for 2 weeks and formed colonies were stained with crystal violet. Data depicted as mean + s.e.m. from one representative experiment performed in triplicate.
Mentions: To evaluate the effects of API5 on cell proliferation, the API5 expression vector or the control vector (no insert) were transfected into CaSki cells which have a low level of API5 expression. Non-tumorigenic HEK 293 cells with low background of API5 expression level were used as a positive control for comparison. Conversely, siRNA targeting API5 (siAPI5) or GFP (irrelevant negative control, siGFP) were also transfected into HeLa cells which have a high level of API5 expression. API5 expression level in the transfected cells were detected by western blotting (Figure 2A). The cell growth assay revealed that cell growth rate in both of the API5-transfected cells was significantly higher than control groups (Figure 2B). Similar increase was also observed in colony formation assay (Figure 2C). In contrast, knock-down of API5 in HeLa cells significantly decreased both of the cell growth rate and colony formation efficacy compared with siGFP control group (Figure 2B and C). These data demonstrate that API5 has a key role in cell proliferation and colony formation of cervical cancer cells.Figure 2

Bottom Line: Immunohistochemical staining showed that API5 expression increased during the normal to tumor transition of cervical carcinoma (P < 0.001), and this increased expression was significantly associated with tumor stage (P = 0.004), tumor grade (P < 0.001), and chemo-radiation response (P = 0.004).In multivariate analysis, API5+ (P = 0.039) and combined API5+/pERK1/2+ (P = 0.032) were independent prognostic factors for overall survival.API5 expression is associated with pERK1/2 in a subset of cervical cancer patients and its expression predicts poor overall survival, supporting that API5 may be a promising novel target for therapeutic interventions.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Gangnam Severance Hospital, Yonsei University College of Medicine, 146-92 Dogok-Dong, Gangnam-Gu, Seoul 135-720, South Korea. genejock@helix.nih.gov.

ABSTRACT

Background: The apoptosis inhibitor-5 (API5), anti-apoptosis protein, is considered a key molecule in the tumor progression and malignant phenotype of tumor cells. Here, we investigated API5 expression in cervical cancer, its clinical significance, and its relationship with phosphorylated extracellular signal-regulated kinase 1 and 2 (pERK1/2) in development and progression of cervical cancer.

Methods: API5 effects on cell growth were assessed in cervical cancer cell lines. API5 and pERK1/2 immunohistochemical staining were performed on a cervical cancer tissue microarray consisting of 173 primary cervical cancers, 306 cervical intraepithelial neoplasias (CINs), and 429 matched normal tissues.

Results: API5 overexpression promoted cell proliferation and colony formation in CaSki cells, whereas API5 knockdown inhibited the both properties in HeLa cells. Immunohistochemical staining showed that API5 expression increased during the normal to tumor transition of cervical carcinoma (P < 0.001), and this increased expression was significantly associated with tumor stage (P = 0.004), tumor grade (P < 0.001), and chemo-radiation response (P = 0.004). API5 expression levels were positively associated with pERK1/2 in cervical cancer (P < 0.001) and high grade CIN (P = 0.031). In multivariate analysis, API5+ (P = 0.039) and combined API5+/pERK1/2+ (P = 0.032) were independent prognostic factors for overall survival.

Conclusions: API5 expression is associated with pERK1/2 in a subset of cervical cancer patients and its expression predicts poor overall survival, supporting that API5 may be a promising novel target for therapeutic interventions.

Show MeSH
Related in: MedlinePlus