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Overexpression of UbcH10 alternates the cell cycle profile and accelerate the tumor proliferation in colon cancer.

Fujita T, Ikeda H, Taira N, Hatoh S, Naito M, Doihara H - BMC Cancer (2009)

Bottom Line: UbcH10 participates in proper metaphase to anaphase transition, and abrogation of UbcH10 results in the premature separation of sister chromatids.Immunohistochemical analysis indicated that UbcH10 was significantly higher in colon cancer tissue compared with normal colon epithelia.Furthermore, the clinicopathological evaluation revealed that UbcH10 was associated with high-grade histological tumors.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cancer and Thoracic Surgery, Okayama University School of Medicine, Okayama, Japan. cqs03255@nifty.com

ABSTRACT

Background: UbcH10 participates in proper metaphase to anaphase transition, and abrogation of UbcH10 results in the premature separation of sister chromatids. To assess the potential role of UbcH10 in colon cancer progression, we analyzed the clinicopathological relevance of UbcH10 in colon cancer.

Methods: We firstly screened the expression profile of UbcH10 in various types of cancer tissues as well as cell lines. Thereafter, using the colon cancer cells line, we manipulated the expression of UbcH10 and evaluated the cell cycle profile and cellular proliferations. Furthermore, the clinicopathological significance of UbcH10 was immunohistologically evaluated in patients with colon cancer. Statistical analysis was performed using the student's t-test and Chi-square test.

Results: Using the colon cancer cells, depletion of UbcH10 resulted in suppression of cellular growth whereas overexpression of UbcH10 promoted the cellular growth and oncogenic cellular growth. Mitotic population was markedly alternated by the manipulation of UbcH10 expression. Immunohistochemical analysis indicated that UbcH10 was significantly higher in colon cancer tissue compared with normal colon epithelia. Furthermore, the clinicopathological evaluation revealed that UbcH10 was associated with high-grade histological tumors.

Conclusion: The results show the clinicopathological significance of UbcH10 in the progression of colon cancer. Thus UbcH10 may act as a novel biomarker in patients with colon cancer.

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Overexpression of UbcH10 promotes the cellular proliferation and alters the cell cycle profile in colon cancer. (A) UbcH10 overexpression in DLD1 colon cancer cells. (B) Cellular proliferation was markedly increased in UbcH10-overexpressed DLD1 cells compared with control cells. (C) Results of anchorage-independent colony formation soft-agar assay. The number of colonies was markedly increased in stably-transfected UbcH10 DLD1 colon cancer cells. (D) Cell cycle profile of overexpresed UbcH10 and control DLD1 cells shows that the population of G2/M cells is moderately decreased with the delivery of UbcH10. (E) a. Immunofluorescence with phosphorylated-histone H3 (pH3). The population of pH3 positive cells was decreased in UbcH10-overexpressed DLD1 cells. b. Quantification of immunofluorescence analysis (triplicate experiments).
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Figure 2: Overexpression of UbcH10 promotes the cellular proliferation and alters the cell cycle profile in colon cancer. (A) UbcH10 overexpression in DLD1 colon cancer cells. (B) Cellular proliferation was markedly increased in UbcH10-overexpressed DLD1 cells compared with control cells. (C) Results of anchorage-independent colony formation soft-agar assay. The number of colonies was markedly increased in stably-transfected UbcH10 DLD1 colon cancer cells. (D) Cell cycle profile of overexpresed UbcH10 and control DLD1 cells shows that the population of G2/M cells is moderately decreased with the delivery of UbcH10. (E) a. Immunofluorescence with phosphorylated-histone H3 (pH3). The population of pH3 positive cells was decreased in UbcH10-overexpressed DLD1 cells. b. Quantification of immunofluorescence analysis (triplicate experiments).

Mentions: To analyze the role of UbcH10 in promoting cellular growth associated with cell cycle progression through mitosis, we overexpressed UbcH10 in colon cancer cells (Fig. 2A). As shown in Fig. 2B, UbcH10 overexpression in DLD1 cells resulted in a significant acceleration of cellular growth. There are remarkable changes of doubling times by the overexpression of UbcH10. Calculated doubling times of DLD1 cells; 37.0 hrs (control cells), 21.5 hrs (UbcH10-overexpressed cells), respectively. To test the effect of UbcH10 on cell growth and oncogenic colony formation, we conducted an anchorage-independent growth assay and further measured the cell cycle profile in DLD1 cells. UbcH10 overexpression promoted an increase in the number and size (data not shown) of colonies on soft agar (Fig. 2C). Recent studies have demonstrated that UbcH10 promotes the metaphase to anaphase transition. To validate the role of UbcH10 in cell cycle regulation, we examined the cell cycle profile in UbcH10 overexpressed colon cancer cells. UbcH10 overexpression in DLD1 cells reduced the G2/M-cell fraction (Fig. 2D). To determine whether a decrease in the G2/M fraction was dependant on the G2 or M phase, we analyzed the mitotic population with immunofluorescence using phosphorylated-histone H3. As shown in Fig. 2E, the percentage of mitotic cells was markedly reduced in UbcH10-abundant DLD1 cells compared with control cells. These results suggest that increases of UbcH10 protein levels could lead to aberrant cell cycle progression, particularly during mitosis, which, in turn, could promote oncogenesis in colon cancer cells.


Overexpression of UbcH10 alternates the cell cycle profile and accelerate the tumor proliferation in colon cancer.

Fujita T, Ikeda H, Taira N, Hatoh S, Naito M, Doihara H - BMC Cancer (2009)

Overexpression of UbcH10 promotes the cellular proliferation and alters the cell cycle profile in colon cancer. (A) UbcH10 overexpression in DLD1 colon cancer cells. (B) Cellular proliferation was markedly increased in UbcH10-overexpressed DLD1 cells compared with control cells. (C) Results of anchorage-independent colony formation soft-agar assay. The number of colonies was markedly increased in stably-transfected UbcH10 DLD1 colon cancer cells. (D) Cell cycle profile of overexpresed UbcH10 and control DLD1 cells shows that the population of G2/M cells is moderately decreased with the delivery of UbcH10. (E) a. Immunofluorescence with phosphorylated-histone H3 (pH3). The population of pH3 positive cells was decreased in UbcH10-overexpressed DLD1 cells. b. Quantification of immunofluorescence analysis (triplicate experiments).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2666760&req=5

Figure 2: Overexpression of UbcH10 promotes the cellular proliferation and alters the cell cycle profile in colon cancer. (A) UbcH10 overexpression in DLD1 colon cancer cells. (B) Cellular proliferation was markedly increased in UbcH10-overexpressed DLD1 cells compared with control cells. (C) Results of anchorage-independent colony formation soft-agar assay. The number of colonies was markedly increased in stably-transfected UbcH10 DLD1 colon cancer cells. (D) Cell cycle profile of overexpresed UbcH10 and control DLD1 cells shows that the population of G2/M cells is moderately decreased with the delivery of UbcH10. (E) a. Immunofluorescence with phosphorylated-histone H3 (pH3). The population of pH3 positive cells was decreased in UbcH10-overexpressed DLD1 cells. b. Quantification of immunofluorescence analysis (triplicate experiments).
Mentions: To analyze the role of UbcH10 in promoting cellular growth associated with cell cycle progression through mitosis, we overexpressed UbcH10 in colon cancer cells (Fig. 2A). As shown in Fig. 2B, UbcH10 overexpression in DLD1 cells resulted in a significant acceleration of cellular growth. There are remarkable changes of doubling times by the overexpression of UbcH10. Calculated doubling times of DLD1 cells; 37.0 hrs (control cells), 21.5 hrs (UbcH10-overexpressed cells), respectively. To test the effect of UbcH10 on cell growth and oncogenic colony formation, we conducted an anchorage-independent growth assay and further measured the cell cycle profile in DLD1 cells. UbcH10 overexpression promoted an increase in the number and size (data not shown) of colonies on soft agar (Fig. 2C). Recent studies have demonstrated that UbcH10 promotes the metaphase to anaphase transition. To validate the role of UbcH10 in cell cycle regulation, we examined the cell cycle profile in UbcH10 overexpressed colon cancer cells. UbcH10 overexpression in DLD1 cells reduced the G2/M-cell fraction (Fig. 2D). To determine whether a decrease in the G2/M fraction was dependant on the G2 or M phase, we analyzed the mitotic population with immunofluorescence using phosphorylated-histone H3. As shown in Fig. 2E, the percentage of mitotic cells was markedly reduced in UbcH10-abundant DLD1 cells compared with control cells. These results suggest that increases of UbcH10 protein levels could lead to aberrant cell cycle progression, particularly during mitosis, which, in turn, could promote oncogenesis in colon cancer cells.

Bottom Line: UbcH10 participates in proper metaphase to anaphase transition, and abrogation of UbcH10 results in the premature separation of sister chromatids.Immunohistochemical analysis indicated that UbcH10 was significantly higher in colon cancer tissue compared with normal colon epithelia.Furthermore, the clinicopathological evaluation revealed that UbcH10 was associated with high-grade histological tumors.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cancer and Thoracic Surgery, Okayama University School of Medicine, Okayama, Japan. cqs03255@nifty.com

ABSTRACT

Background: UbcH10 participates in proper metaphase to anaphase transition, and abrogation of UbcH10 results in the premature separation of sister chromatids. To assess the potential role of UbcH10 in colon cancer progression, we analyzed the clinicopathological relevance of UbcH10 in colon cancer.

Methods: We firstly screened the expression profile of UbcH10 in various types of cancer tissues as well as cell lines. Thereafter, using the colon cancer cells line, we manipulated the expression of UbcH10 and evaluated the cell cycle profile and cellular proliferations. Furthermore, the clinicopathological significance of UbcH10 was immunohistologically evaluated in patients with colon cancer. Statistical analysis was performed using the student's t-test and Chi-square test.

Results: Using the colon cancer cells, depletion of UbcH10 resulted in suppression of cellular growth whereas overexpression of UbcH10 promoted the cellular growth and oncogenic cellular growth. Mitotic population was markedly alternated by the manipulation of UbcH10 expression. Immunohistochemical analysis indicated that UbcH10 was significantly higher in colon cancer tissue compared with normal colon epithelia. Furthermore, the clinicopathological evaluation revealed that UbcH10 was associated with high-grade histological tumors.

Conclusion: The results show the clinicopathological significance of UbcH10 in the progression of colon cancer. Thus UbcH10 may act as a novel biomarker in patients with colon cancer.

Show MeSH
Related in: MedlinePlus