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KIAA0101 (OEACT-1), an expressionally down-regulated and growth-inhibitory gene in human hepatocellular carcinoma.

Guo M, Li J, Wan D, Gu J - BMC Cancer (2006)

Bottom Line: The in vitro cell growth curve was used for examining the effect of over-expression of KIAA0101 in HCC cells.Western blot showed KIAA0101 protein expression was down-regulated in HCC tissues as compared with their counterpart non-cancerous liver tissues in 25 out of 30 cases.This gene could inhibit the HCC cell growth in vitro and presumably by its blocking effect on cell cycle.

View Article: PubMed Central - HTML - PubMed

Affiliation: Shanghai Medical College, Fudan University, Shanghai, China. guomingleicn@yahoo.com.cn

ABSTRACT

Background: Our previous cDNA array results indicated KIAA0101 as one of the differentially expressed genes in human hepatocellular carcinoma (HCC) as compared with non-cancerous liver. However, it is necessary to study its expression at protein level in HCC and its biological function for HCC cell growth.

Method: Western blot and tissue array were performed to compare KIAA0101 protein expression level in paired human HCC and non-cancerous liver tissues from the same patients. Investigation of its subcellular localization was done by using dual fluorescence image examination and enriched mitochondrial protein Western blot analysis. The in vitro cell growth curve was used for examining the effect of over-expression of KIAA0101 in HCC cells. FACS was used to analyze the cell cycle pattern in KIAA0101 expression positive (+) and negative (-) cell populations isolated by the pMACSKKII system after KIAA0101 cDNA transfection.

Results: Western blot showed KIAA0101 protein expression was down-regulated in HCC tissues as compared with their counterpart non-cancerous liver tissues in 25 out of 30 cases. Tissue array also demonstrated the same pattern in 161 paired samples. KIAA0101 was predominantly localized in mitochondria and partially in nuclei. KIAA0101 cDNA transfection could inhibit the HCC cell growth in vitro. In cell cycle analysis, it could arrest cells at the G1 to S phase transition.

Conclusion: KIAA0101 protein expression was down-regulated in HCC. This gene could inhibit the HCC cell growth in vitro and presumably by its blocking effect on cell cycle.

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KIAA 0101 expression in tissue array was analyzed by immunohistochemistry (ISH) examination. The tissue array showed different levels of KIAA0101 protein expression in paired HCC and non-cancerous tissues, liver cirrhosis and normal liver tissues by ISH method. Weak expression of KIAA0101 was observed in HCC; while strong expression in its counterpart non-cancerous liver tissue. In 8 out of 13 liver cirrhosis and all 10 normal liver tissues, KIAA0101 protein expression level were strong(+++) or moderate(++) level.
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Figure 3: KIAA 0101 expression in tissue array was analyzed by immunohistochemistry (ISH) examination. The tissue array showed different levels of KIAA0101 protein expression in paired HCC and non-cancerous tissues, liver cirrhosis and normal liver tissues by ISH method. Weak expression of KIAA0101 was observed in HCC; while strong expression in its counterpart non-cancerous liver tissue. In 8 out of 13 liver cirrhosis and all 10 normal liver tissues, KIAA0101 protein expression level were strong(+++) or moderate(++) level.

Mentions: To demonstrate KIAA0101 expression pattern at the protein level in human HCC, 30 human HCC (T) and matched noncancerous liver tissues (NT) from the same patients were analyzed by Western blot (Figure 2). The protein band intensity was compared after being normalized with that of the beta-actin internal control. It was striking that the protein expression level of KIAA0101 in HCC tissues was reduced as compared with that of non-cancerous tissues among 25 out of 30 paired samples. We have conducted the tissue array by immunohistochemical survey(ISH) for further validation. Fig 3 illustrated the different intensity between HCC and matched non-cancerous tissues, liver cirrhosis and normal liver tissues by ISH method. As demonstrated in table 1 (see Additional file 1), the tissue array showed the signal intensity was weak(+) or negative(-) in 107 out of 161 HCC tissues; while the weak (+) or negative (-) signals were seen in only 24 out of 161 samples of non-cancerous liver tissues. Therefore, the KIAA0101 protein expression was much lower in the HCC tissues as compared with the non-cancerous tissues (p < 0.01). In addition, in 8 out of 13 of liver cirrhosis tissues and all 10 normal liver tissues, KIAA0101 protein was expressed varying from moderate(++) to high(+++) level. However, the KIAA0101 protein level in HCC tissues was obviously reduced. Furthermore, we have attempted to investigate if the down-regulation of KIAA0101 protein expression had any correlation with the patients' background information, such as age, sex, histopathological grading, tumor size and HBV infection. However, we have not found any significant association between the clinical or pathological state of HCC with the KIAA0101 expression (see Additional file 2).


KIAA0101 (OEACT-1), an expressionally down-regulated and growth-inhibitory gene in human hepatocellular carcinoma.

Guo M, Li J, Wan D, Gu J - BMC Cancer (2006)

KIAA 0101 expression in tissue array was analyzed by immunohistochemistry (ISH) examination. The tissue array showed different levels of KIAA0101 protein expression in paired HCC and non-cancerous tissues, liver cirrhosis and normal liver tissues by ISH method. Weak expression of KIAA0101 was observed in HCC; while strong expression in its counterpart non-cancerous liver tissue. In 8 out of 13 liver cirrhosis and all 10 normal liver tissues, KIAA0101 protein expression level were strong(+++) or moderate(++) level.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1483895&req=5

Figure 3: KIAA 0101 expression in tissue array was analyzed by immunohistochemistry (ISH) examination. The tissue array showed different levels of KIAA0101 protein expression in paired HCC and non-cancerous tissues, liver cirrhosis and normal liver tissues by ISH method. Weak expression of KIAA0101 was observed in HCC; while strong expression in its counterpart non-cancerous liver tissue. In 8 out of 13 liver cirrhosis and all 10 normal liver tissues, KIAA0101 protein expression level were strong(+++) or moderate(++) level.
Mentions: To demonstrate KIAA0101 expression pattern at the protein level in human HCC, 30 human HCC (T) and matched noncancerous liver tissues (NT) from the same patients were analyzed by Western blot (Figure 2). The protein band intensity was compared after being normalized with that of the beta-actin internal control. It was striking that the protein expression level of KIAA0101 in HCC tissues was reduced as compared with that of non-cancerous tissues among 25 out of 30 paired samples. We have conducted the tissue array by immunohistochemical survey(ISH) for further validation. Fig 3 illustrated the different intensity between HCC and matched non-cancerous tissues, liver cirrhosis and normal liver tissues by ISH method. As demonstrated in table 1 (see Additional file 1), the tissue array showed the signal intensity was weak(+) or negative(-) in 107 out of 161 HCC tissues; while the weak (+) or negative (-) signals were seen in only 24 out of 161 samples of non-cancerous liver tissues. Therefore, the KIAA0101 protein expression was much lower in the HCC tissues as compared with the non-cancerous tissues (p < 0.01). In addition, in 8 out of 13 of liver cirrhosis tissues and all 10 normal liver tissues, KIAA0101 protein was expressed varying from moderate(++) to high(+++) level. However, the KIAA0101 protein level in HCC tissues was obviously reduced. Furthermore, we have attempted to investigate if the down-regulation of KIAA0101 protein expression had any correlation with the patients' background information, such as age, sex, histopathological grading, tumor size and HBV infection. However, we have not found any significant association between the clinical or pathological state of HCC with the KIAA0101 expression (see Additional file 2).

Bottom Line: The in vitro cell growth curve was used for examining the effect of over-expression of KIAA0101 in HCC cells.Western blot showed KIAA0101 protein expression was down-regulated in HCC tissues as compared with their counterpart non-cancerous liver tissues in 25 out of 30 cases.This gene could inhibit the HCC cell growth in vitro and presumably by its blocking effect on cell cycle.

View Article: PubMed Central - HTML - PubMed

Affiliation: Shanghai Medical College, Fudan University, Shanghai, China. guomingleicn@yahoo.com.cn

ABSTRACT

Background: Our previous cDNA array results indicated KIAA0101 as one of the differentially expressed genes in human hepatocellular carcinoma (HCC) as compared with non-cancerous liver. However, it is necessary to study its expression at protein level in HCC and its biological function for HCC cell growth.

Method: Western blot and tissue array were performed to compare KIAA0101 protein expression level in paired human HCC and non-cancerous liver tissues from the same patients. Investigation of its subcellular localization was done by using dual fluorescence image examination and enriched mitochondrial protein Western blot analysis. The in vitro cell growth curve was used for examining the effect of over-expression of KIAA0101 in HCC cells. FACS was used to analyze the cell cycle pattern in KIAA0101 expression positive (+) and negative (-) cell populations isolated by the pMACSKKII system after KIAA0101 cDNA transfection.

Results: Western blot showed KIAA0101 protein expression was down-regulated in HCC tissues as compared with their counterpart non-cancerous liver tissues in 25 out of 30 cases. Tissue array also demonstrated the same pattern in 161 paired samples. KIAA0101 was predominantly localized in mitochondria and partially in nuclei. KIAA0101 cDNA transfection could inhibit the HCC cell growth in vitro. In cell cycle analysis, it could arrest cells at the G1 to S phase transition.

Conclusion: KIAA0101 protein expression was down-regulated in HCC. This gene could inhibit the HCC cell growth in vitro and presumably by its blocking effect on cell cycle.

Show MeSH
Related in: MedlinePlus