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Objective After the detection of FAT10 protein in human cervical cancer tissue behind clinical pathological examination,investigate the effect of up regulation and down regulating the expression of FAT10 gene on the radiation sensitivity of cervical cancer cell line SiHa cells in nude mice.Methods The expression levels of FAT10 mRNA in 30 cases cervical cancer and adjacent tissues were detected by RT-PCR,FAT10 siRNA and recombinant expression vector pcDNA3.1-FAT10 were respectively transfected into cervical cancer SiHa cells by electroporation,and stable transfection cell lines was obtained by G418 screening.After transfection 48h,the expression levels of FAT10 mRNA and protein were respectively detected by fluorescence quantitative RT-PCR and Western blot.Cell proliferation was detected with CCK-8.The effect of FAT10 expression on the radiation sensitivity of cervical cancer cell SiHa line was observed by the clonogenic experiment.The cell apoptosis was detected by TUNEL.The growth of cervical cancer cells was detected by Xenograft tumor assay combined with X-ray irradiation.Results The relative expression level of FAT10 protein in human cervical cancer tissues was significantly higher than that in adjacent tissues,and the difference was statistically significant(P ＜ 0.05).The expression of FAT10 gene in cervical cancer SiHa cells was significantly lower in FAT10 down group,and the FAT10 up regulation group was significantly higher.The radiation sensitivity of cervical cancer SiHa cells was increased in the FAT10 down group,and the radiation sensitivity of SiHa cells was decreased in the FAT10 up regulation group (P ＜ 0.05).FAT10 down group of cervical cancer SiHa cell proliferation was inhibited,the cell apoptosis rate was increased (P ＜ 0.05).The proliferation ability of FAT10 cells was significantly enhanced,and the cell apoptosis rate was decreased in the FAT10 up regulation group (P ＜ 0.05).The average volume of transplanted tumor in the FAT10 down group was significantly smaller than that of the control group (P ＜ 0.05);the average volume of the tumor in the FAT10 up regulation group was larger than that of the control group (P ＜ 0.05).The average volume of the tumor was significantly decreased in the FAT10 down group after irradiation of 20Gy gamma ray(P ＜ 0.05),and the average volume of subcutaneous implanted tumor in nude mice was not significantly different (P ＞ 0.05).Conclusion Downregulation of FAT10 expression can inhibit the growth of human cervical cancer SiHa cells in nude mice,enhance the radiation sensitivity of tumor cells,up regulation of FAT10 expression will make tumor radiation resistance.
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Objective:To investigate the correlation of FAT10 expression with the malignant characteristics of hepatocellular car-cinoma (HCC), and to explore the effect of FAT10 on RhoA and cytoskeleton of HCC. Methods: Immunohistochemistry (IHC) was used to detect the FAT10 expression level of 108 HCC patients, and the correlation between the expression of FAT10 and the malignant characteristics of HCC patients was analyzed. We transiently transfected plasmids with overexpressed FAT10 using 7721 and HepG2 cells or interfered with FAT10 expression using siRNA in Huh7 and LM3 cells. Active-RhoA, total-RhoA, and ROCK protein expres-sion levels were detected by Western blot analysis after overexpression or interference. We also used immunofluorescence to detect changes in the cytoskeleton protein F-actin after FAT10 overexpression in 7721 cells. Results:Correlation analysis showed that both ac-tive-RhoA and FAT10 expression levels were significantly correlated with clinical malignant characteristics by using IHC (RhoA:me-tastasis, P=0.036 and recurrence, P=0.026;FAT10:metastasis, P=0.031 and recurrence P=0.004). In addition, active-RhoA expression level was correlated with FAT10 (P=0.000). Survival analysis showed that the prognoses of low-expression active RhoA (P=0.019) or FAT10 (P=0.026) groups were significantly better than those of the high-expression groups. Western blot analysis showed that FAT10 increased the expression of active-RhoA and ROCK. However, the expression of active-RhoA and ROCK decreased after FAT10 inter-ference. F-actin expression increased in the 7721 cells with overexpressed FAT10 (all P<0.01). Moreover, FAT10 facilitated F-actin ag-gregation on cell membrane and changes in F-actin. Conclusion:FAT10 is correlated with the malignant characteristics of HCC and may promote changes in HCC cytoskeleton induced by active-RhoA.
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Objective: To study the relationship between FAT10 expression and biological behaviors in inif trating ductal carcinoma of breast. Methods: The expressions of diubiquitin (FAT10), estrogen receptor (ER), progesterone receptor (PR) and c-erbB2 in 50 cases of inif trating ductal carcinoma of breast were detected by immunohistochemistry. Western blot was used to detect FAT10 expression in MB-MDA-435, MB-MDA-435-transfected with FAT10 siRNA expression plasmid, MCF-7 and MCF-7-transfected with FAT10 expression plasmid, respectively. Transwell was used to detect invasion capability of MB-MDA-435, MB-MDA-435-transfected with FAT10 siRNA expression plasmid, MCF-7 and MCF-7-transfected with FAT10 expression plasmid. Results: hTe expression intensity of FAT10 was signiifcantly correlated to patho-grading, lymph nodes metastasis, distant metastasis and TNM staging (P0.05). hTe expression intensity of FAT10 in receptor- negative group was obviously stronger than that in receptor- positive group (P<0.01). hTe expression intensity of FAT10 in triple-negative breast cancer was signiifcantly stronger than that in non- triple-negative breast cancer (P<0.01). hTe survival rate of patients with FAT10 positive expression was significantly lower than negative ones (P<0.05). Western blot results showed that FAT10 intensity in MB-MDA-435 significantly higher than that in MCF-7. Up-regulation expression of FAT10 could obviously increase the invasion capability of MCF-7, and down-regulation of FAT10 could signiifcantly decrease the invasion capability of MB-MDA-435 (P<0.01). Conclusion: FAT10 might increase the invasion capability of breast cancer cells by direct or indirect ways, and play an important role in invasion and metastasis of breast cancer. FAT10 might be an independent index for evaluation of breast cancer prognosis, and a potential target for breast cancer therapy, especially for triple-negative breast cancer.
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Ubiquitin-like proteins are structurally similar to ubiquitin. These proteins are processed, activated, conjugated, and re-leased from conjugates by enzymatic steps that are similar to the corresponding mechanisms for ubiquitin. Ubiquitin-like proteins regu-late a wide array of cellular processes through modification processes, such as nuclear-cytosolic transport, transcriptional regulation, protein stability, response to stress, and progression through the cell cycle. A large number of recent studies have found dysfunctional ubiquitin-like proteins in hepatocellular carcinoma. These proteins are important in tumorigenesis, cell proliferation, apoptosis, and an-giogenesis. Anticancer drug studies revealed that regulating protein modification by using ubiquitin-like proteins may alter the anti-tu-mor effects of chemotherapy and thus influence the chemosensitivity of hepatocellular carcinoma. Results indicate that ubiquitin-like proteins may become a new target for cancer therapy. The mechanism of ubiquitin-like proteins in tumorigenesis and hepatocellular car-cinoma progression is of great significance in the diagnosis and treatment of hepatocellular carcinoma.
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Objective To investigate the expressions of FAT10 and p53 mutant in gastric cancer tissues and their relations. Methods Immunohistochemistry and RT-PCR were used to detect the expressions of FAT10 and p53 in gastric cancer tissues (n=62), para-cancerous tissues (2-5 cm apart from cancer, n=62), and normal gastric tissues (7>5 cm apart from cancer, n=62). The association of FAT10 with p53 and clinical outcomes were analyzed by Spearman and Pearson correlation. Results The immunohistochemistry examination showed that expressions of FAT10 [51.61%(32/62)] and p53 [45.16% (28/62)] were significantly higher in cancerous tissues than in para-cancerous tissues [12.90%(8/62) and 14.51% (9/62), χ2=21.26 and 20.69, P<0.01] and normal tissues [6.45% (4/62) and 9.68% (6/62), χ2=13.91 and 19.61, P<0.01]. Overexpressions of FAT10 protein and mRNA in cancerous tissues were closely related to lymph node metastasis and TNM staging (both P value<0.05). There was a positive correlation between FAT10 and p53 in protein and mRNA expressions (protein r=0. 865, P<0.05; mRNA r=0.761, P< 0.01). Those with positive expression of FAT10 had lower survival rate compared to those with negative expression (P<0.05). Conclusions The positive relation between over-expression of FAT10 and p53 implicates that both are involved in the gastric carcinogenesis, and FAT10 is a novel gastric cancer marker with prognostic significance.