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Actin-like 6A predicts poor prognosis of hepatocellular carcinoma and promotes metastasis and epithelial-mesenchymal transition.

Xiao S, Chang RM, Yang MY, Lei X, Liu X, Gao WB, Xiao JL, Yang LY - Hepatology (2016)

Bottom Line: Opposite results are observed when ACTL6A is knocked down.ACTL6A knockdown has the equal blockage effect as the Notch signaling inhibitor, N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butylester, in HCC cells.Further studies indicate that ACTL6A might manipulate SRY (sex determining region Y)-box 2 (SOX2) expression and then activate Notch1 signaling.

View Article: PubMed Central - PubMed

Affiliation: Liver Cancer Laboratory, Department of Surgery, Xiangya Hospital, Central South University, Changsha, China.

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ACTL6A promotes proliferation and metastasis of HCC in vitro and in vivo. (A) Proliferation of PLC/PRF5‐ACTL6A, Hep3B‐shACTL6A‐3 cells and control cells was examined by MTT and colony formation assays (B). (C) Wound‐healing assay and (D) transwell invasion assay were subjected to detect the migration and invasion capacity of ACTL6A‐interfered cells. (E) SC tumors from PLC/PRF5‐ACTL6A and Hep3B‐shACTL6A cells and their control cells are shown in the upper two panels. Orthotopic tumors from each indicated groups are shown in the lower two panels (black arrows indicate tumors). Tumor volumes of tumors are shown in the right panels. (F) Representative pictures of intrahepatic and lung metastasis; metastatic nodules proportion of livers or lungs was calculated and compared.*P < 0.05; **P < 0.01 based on the Student t test. Error bars, standard deviation.
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hep28417-fig-0002: ACTL6A promotes proliferation and metastasis of HCC in vitro and in vivo. (A) Proliferation of PLC/PRF5‐ACTL6A, Hep3B‐shACTL6A‐3 cells and control cells was examined by MTT and colony formation assays (B). (C) Wound‐healing assay and (D) transwell invasion assay were subjected to detect the migration and invasion capacity of ACTL6A‐interfered cells. (E) SC tumors from PLC/PRF5‐ACTL6A and Hep3B‐shACTL6A cells and their control cells are shown in the upper two panels. Orthotopic tumors from each indicated groups are shown in the lower two panels (black arrows indicate tumors). Tumor volumes of tumors are shown in the right panels. (F) Representative pictures of intrahepatic and lung metastasis; metastatic nodules proportion of livers or lungs was calculated and compared.*P < 0.05; **P < 0.01 based on the Student t test. Error bars, standard deviation.

Mentions: To understand the function of ACTL6A in HCC cells, we manipulated ACTL6A expression in cells by ectopic expression and short hairpin RNA (shRNA) knockdown. Ectopic ACTL6A was constituently expressed in PLC/PRF5 cells named as PLC/PRF5‐ACTL6A subsequently. Meanwhile, three shRNAs (shRNA1, shRNA2, and shRNA3) were designed to silence ACTL6A expression in Hep3B cells named as Hep3B‐shACTL6A subsequently. Expression level of ACTL6A was identified by real‐time PCR and western blotting; shRNA3 was the most effective one and was chosen for further study (Supporting Fig. 4A,B). Compared to PLC/PRF5, PLC/PRF5‐ACTL6A had a higher absorbance in methyl thiazol tetrazolium assay, which indicated a higher proliferation rate (Fig. 2A). Consistently, PLC/PRF5‐ACTL6A cells also formed more colonies in colony formation assay (Fig. 2B and Supporting Fig. 4C). In contrast, Hep3B‐shACTL6A cells had decreased absorbance, cell proliferation rate, and clonogenicity capacity (Fig. 2A,B and Supporting Fig. 4C). The wound‐healing and transwell assays were used to investigate migration and invasion capacity. Results showed that PLC/PRF5‐ACTL6A cells had a faster wound closure rate and more invasion cells than PLC/PRF5 cells, whereas Hep3B‐shACTL6A cells had markedly reduced migratory and invasive capacity (Fig. 2C,D and Supporting Fig. 4D,E). It suggests that ACTL6A promotes HCC cell proliferation, migration, and invasion capacity in vitro.


Actin-like 6A predicts poor prognosis of hepatocellular carcinoma and promotes metastasis and epithelial-mesenchymal transition.

Xiao S, Chang RM, Yang MY, Lei X, Liu X, Gao WB, Xiao JL, Yang LY - Hepatology (2016)

ACTL6A promotes proliferation and metastasis of HCC in vitro and in vivo. (A) Proliferation of PLC/PRF5‐ACTL6A, Hep3B‐shACTL6A‐3 cells and control cells was examined by MTT and colony formation assays (B). (C) Wound‐healing assay and (D) transwell invasion assay were subjected to detect the migration and invasion capacity of ACTL6A‐interfered cells. (E) SC tumors from PLC/PRF5‐ACTL6A and Hep3B‐shACTL6A cells and their control cells are shown in the upper two panels. Orthotopic tumors from each indicated groups are shown in the lower two panels (black arrows indicate tumors). Tumor volumes of tumors are shown in the right panels. (F) Representative pictures of intrahepatic and lung metastasis; metastatic nodules proportion of livers or lungs was calculated and compared.*P < 0.05; **P < 0.01 based on the Student t test. Error bars, standard deviation.
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hep28417-fig-0002: ACTL6A promotes proliferation and metastasis of HCC in vitro and in vivo. (A) Proliferation of PLC/PRF5‐ACTL6A, Hep3B‐shACTL6A‐3 cells and control cells was examined by MTT and colony formation assays (B). (C) Wound‐healing assay and (D) transwell invasion assay were subjected to detect the migration and invasion capacity of ACTL6A‐interfered cells. (E) SC tumors from PLC/PRF5‐ACTL6A and Hep3B‐shACTL6A cells and their control cells are shown in the upper two panels. Orthotopic tumors from each indicated groups are shown in the lower two panels (black arrows indicate tumors). Tumor volumes of tumors are shown in the right panels. (F) Representative pictures of intrahepatic and lung metastasis; metastatic nodules proportion of livers or lungs was calculated and compared.*P < 0.05; **P < 0.01 based on the Student t test. Error bars, standard deviation.
Mentions: To understand the function of ACTL6A in HCC cells, we manipulated ACTL6A expression in cells by ectopic expression and short hairpin RNA (shRNA) knockdown. Ectopic ACTL6A was constituently expressed in PLC/PRF5 cells named as PLC/PRF5‐ACTL6A subsequently. Meanwhile, three shRNAs (shRNA1, shRNA2, and shRNA3) were designed to silence ACTL6A expression in Hep3B cells named as Hep3B‐shACTL6A subsequently. Expression level of ACTL6A was identified by real‐time PCR and western blotting; shRNA3 was the most effective one and was chosen for further study (Supporting Fig. 4A,B). Compared to PLC/PRF5, PLC/PRF5‐ACTL6A had a higher absorbance in methyl thiazol tetrazolium assay, which indicated a higher proliferation rate (Fig. 2A). Consistently, PLC/PRF5‐ACTL6A cells also formed more colonies in colony formation assay (Fig. 2B and Supporting Fig. 4C). In contrast, Hep3B‐shACTL6A cells had decreased absorbance, cell proliferation rate, and clonogenicity capacity (Fig. 2A,B and Supporting Fig. 4C). The wound‐healing and transwell assays were used to investigate migration and invasion capacity. Results showed that PLC/PRF5‐ACTL6A cells had a faster wound closure rate and more invasion cells than PLC/PRF5 cells, whereas Hep3B‐shACTL6A cells had markedly reduced migratory and invasive capacity (Fig. 2C,D and Supporting Fig. 4D,E). It suggests that ACTL6A promotes HCC cell proliferation, migration, and invasion capacity in vitro.

Bottom Line: Opposite results are observed when ACTL6A is knocked down.ACTL6A knockdown has the equal blockage effect as the Notch signaling inhibitor, N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butylester, in HCC cells.Further studies indicate that ACTL6A might manipulate SRY (sex determining region Y)-box 2 (SOX2) expression and then activate Notch1 signaling.

View Article: PubMed Central - PubMed

Affiliation: Liver Cancer Laboratory, Department of Surgery, Xiangya Hospital, Central South University, Changsha, China.

Show MeSH
Related in: MedlinePlus