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Functional proteomic analysis reveals the involvement of KIAA1199 in breast cancer growth, motility and invasiveness.

Jami MS, Hou J, Liu M, Varney ML, Hassan H, Dong J, Geng L, Wang J, Yu F, Huang X, Peng H, Fu K, Li Y, Singh RK, Ding SJ - BMC Cancer (2014)

Bottom Line: KIAA1199 mRNA and protein was significantly overexpressed in breast tumor specimens and cell lines as compared with non-neoplastic breast tissues from large-scale microarray and studies of breast cancer cell lines and tumors.The KIAA1199 knockdown cells showed reduced motility and cell proliferation in vitro.Moreover, when the knockdown cells were injected into the mammary fat pads of female athymic nude mice, there was a significant decrease in tumor incidence and growth.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA. rsingh@unmc.edu.

ABSTRACT

Background: KIAA1199 is a recently identified novel gene that is up-regulated in human cancer with poor survival. Our proteomic study on signaling polarity in chemotactic cells revealed KIAA1199 as a novel protein target that may be involved in cellular chemotaxis and motility. In the present study, we examined the functional significance of KIAA1199 expression in breast cancer growth, motility and invasiveness.

Methods: We validated the previous microarray observation by tissue microarray immunohistochemistry using a TMA slide containing 12 breast tumor tissue cores and 12 corresponding normal tissues. We performed the shRNA-mediated knockdown of KIAA1199 in MDA-MB-231 and HS578T cells to study the role of this protein in cell proliferation, migration and apoptosis in vitro. We studied the effects of KIAA1199 knockdown in vivo in two groups of mice (n = 5). We carried out the SILAC LC-MS/MS based proteomic studies on the involvement of KIAA1199 in breast cancer.

Results: KIAA1199 mRNA and protein was significantly overexpressed in breast tumor specimens and cell lines as compared with non-neoplastic breast tissues from large-scale microarray and studies of breast cancer cell lines and tumors. To gain deeper insights into the novel role of KIAA1199 in breast cancer, we modulated KIAA1199 expression using shRNA-mediated knockdown in two breast cancer cell lines (MDA-MB-231 and HS578T), expressing higher levels of KIAA1199. The KIAA1199 knockdown cells showed reduced motility and cell proliferation in vitro. Moreover, when the knockdown cells were injected into the mammary fat pads of female athymic nude mice, there was a significant decrease in tumor incidence and growth. In addition, quantitative proteomic analysis revealed that knockdown of KIAA1199 in breast cancer (MDA-MB-231) cells affected a broad range of cellular functions including apoptosis, metabolism and cell motility.

Conclusions: Our findings indicate that KIAA1199 may play an important role in breast tumor growth and invasiveness, and that it may represent a novel target for biomarker development and a novel therapeutic target for breast cancer.

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KIAA1199 Knockdown inhibited tumorigenicity, growth and neovascularization. A) The relative tumor incidence in MDA-MB-231-ShNC and MDA-MB-231-ShB cell bearing mice; The MDA-MB-231-ShNC (control) and MDA-MB-231-ShB cells were implanted into the mammary fat pads of two groups of nude mice (n = 5). Four of the MDA-MB-231-ShNC and one of the MDA-MB-231-ShB implanted mice developed tumors. B) Tumor growth diagram for MDA-MB-231-ShNC and MDA-MB-231-ShB injected mice.
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Figure 4: KIAA1199 Knockdown inhibited tumorigenicity, growth and neovascularization. A) The relative tumor incidence in MDA-MB-231-ShNC and MDA-MB-231-ShB cell bearing mice; The MDA-MB-231-ShNC (control) and MDA-MB-231-ShB cells were implanted into the mammary fat pads of two groups of nude mice (n = 5). Four of the MDA-MB-231-ShNC and one of the MDA-MB-231-ShB implanted mice developed tumors. B) Tumor growth diagram for MDA-MB-231-ShNC and MDA-MB-231-ShB injected mice.

Mentions: To determine whether KIAA1199 depletion modulates tumor growth, we implanted the MDA-MB-231-ShNC (control) and MDA-MB-231-ShB cells into the mammary fat pads of nude mice (n = 5). We observed significant reduction in tumor incidence following KIAA1199 knockdown (Figure 4A). Four of the MDA-MB-231-ShNC and one of the MDA-MB-231-ShB implanted mice developed tumors. In addition, we observed a significant inhibition in the tumor growth (Figure 4B) in mice bearing the MDA-MB-231-ShB cells as compared to MDA-MB-231-ShNC. We validated the levels of KIAA1199 in tumors using immunohistochemistry. MDA-MB-231-ShNC tumors showed intense KIAA1199 expression whereas MDA-MB-231-ShB tumors showed very little or no immunostaining for KIAA1199 (Figure 5). Moreover, the results showed the cytosolic localization of KIAA1199 protein. Interestingly, several local metastatic foci, expressing even higher levels of KIAA1199, appeared in the fat pads adjacent to the MDA-MB-231-ShNC tumors. These data demonstrate that knockdown of KIAA1199 inhibited MDA-MB-231 tumorigenesis and growth in vivo.


Functional proteomic analysis reveals the involvement of KIAA1199 in breast cancer growth, motility and invasiveness.

Jami MS, Hou J, Liu M, Varney ML, Hassan H, Dong J, Geng L, Wang J, Yu F, Huang X, Peng H, Fu K, Li Y, Singh RK, Ding SJ - BMC Cancer (2014)

KIAA1199 Knockdown inhibited tumorigenicity, growth and neovascularization. A) The relative tumor incidence in MDA-MB-231-ShNC and MDA-MB-231-ShB cell bearing mice; The MDA-MB-231-ShNC (control) and MDA-MB-231-ShB cells were implanted into the mammary fat pads of two groups of nude mice (n = 5). Four of the MDA-MB-231-ShNC and one of the MDA-MB-231-ShB implanted mice developed tumors. B) Tumor growth diagram for MDA-MB-231-ShNC and MDA-MB-231-ShB injected mice.
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Figure 4: KIAA1199 Knockdown inhibited tumorigenicity, growth and neovascularization. A) The relative tumor incidence in MDA-MB-231-ShNC and MDA-MB-231-ShB cell bearing mice; The MDA-MB-231-ShNC (control) and MDA-MB-231-ShB cells were implanted into the mammary fat pads of two groups of nude mice (n = 5). Four of the MDA-MB-231-ShNC and one of the MDA-MB-231-ShB implanted mice developed tumors. B) Tumor growth diagram for MDA-MB-231-ShNC and MDA-MB-231-ShB injected mice.
Mentions: To determine whether KIAA1199 depletion modulates tumor growth, we implanted the MDA-MB-231-ShNC (control) and MDA-MB-231-ShB cells into the mammary fat pads of nude mice (n = 5). We observed significant reduction in tumor incidence following KIAA1199 knockdown (Figure 4A). Four of the MDA-MB-231-ShNC and one of the MDA-MB-231-ShB implanted mice developed tumors. In addition, we observed a significant inhibition in the tumor growth (Figure 4B) in mice bearing the MDA-MB-231-ShB cells as compared to MDA-MB-231-ShNC. We validated the levels of KIAA1199 in tumors using immunohistochemistry. MDA-MB-231-ShNC tumors showed intense KIAA1199 expression whereas MDA-MB-231-ShB tumors showed very little or no immunostaining for KIAA1199 (Figure 5). Moreover, the results showed the cytosolic localization of KIAA1199 protein. Interestingly, several local metastatic foci, expressing even higher levels of KIAA1199, appeared in the fat pads adjacent to the MDA-MB-231-ShNC tumors. These data demonstrate that knockdown of KIAA1199 inhibited MDA-MB-231 tumorigenesis and growth in vivo.

Bottom Line: KIAA1199 mRNA and protein was significantly overexpressed in breast tumor specimens and cell lines as compared with non-neoplastic breast tissues from large-scale microarray and studies of breast cancer cell lines and tumors.The KIAA1199 knockdown cells showed reduced motility and cell proliferation in vitro.Moreover, when the knockdown cells were injected into the mammary fat pads of female athymic nude mice, there was a significant decrease in tumor incidence and growth.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA. rsingh@unmc.edu.

ABSTRACT

Background: KIAA1199 is a recently identified novel gene that is up-regulated in human cancer with poor survival. Our proteomic study on signaling polarity in chemotactic cells revealed KIAA1199 as a novel protein target that may be involved in cellular chemotaxis and motility. In the present study, we examined the functional significance of KIAA1199 expression in breast cancer growth, motility and invasiveness.

Methods: We validated the previous microarray observation by tissue microarray immunohistochemistry using a TMA slide containing 12 breast tumor tissue cores and 12 corresponding normal tissues. We performed the shRNA-mediated knockdown of KIAA1199 in MDA-MB-231 and HS578T cells to study the role of this protein in cell proliferation, migration and apoptosis in vitro. We studied the effects of KIAA1199 knockdown in vivo in two groups of mice (n = 5). We carried out the SILAC LC-MS/MS based proteomic studies on the involvement of KIAA1199 in breast cancer.

Results: KIAA1199 mRNA and protein was significantly overexpressed in breast tumor specimens and cell lines as compared with non-neoplastic breast tissues from large-scale microarray and studies of breast cancer cell lines and tumors. To gain deeper insights into the novel role of KIAA1199 in breast cancer, we modulated KIAA1199 expression using shRNA-mediated knockdown in two breast cancer cell lines (MDA-MB-231 and HS578T), expressing higher levels of KIAA1199. The KIAA1199 knockdown cells showed reduced motility and cell proliferation in vitro. Moreover, when the knockdown cells were injected into the mammary fat pads of female athymic nude mice, there was a significant decrease in tumor incidence and growth. In addition, quantitative proteomic analysis revealed that knockdown of KIAA1199 in breast cancer (MDA-MB-231) cells affected a broad range of cellular functions including apoptosis, metabolism and cell motility.

Conclusions: Our findings indicate that KIAA1199 may play an important role in breast tumor growth and invasiveness, and that it may represent a novel target for biomarker development and a novel therapeutic target for breast cancer.

Show MeSH
Related in: MedlinePlus