Limits...
GNAI1 Suppresses Tumor Cell Migration and Invasion and is Post-Transcriptionally Regulated by Mir-320a/c/d in Hepatocellular Carcinoma.

Yao J, Liang LH, Zhang Y, Ding J, Tian Q, Li JJ, He XH - Cancer Biol Med (2012)

Bottom Line: The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels.GNAI1 is downregulated in HCC and inhibits the migration and invasion of HCC cells.Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China ; State Key Laboratory for Diagnosis and Treatment for Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.

ABSTRACT

Objective: To explore the role and regulation of guanine nucleotide-binding protein G(i), α-1 subunit (GNAI1) in hepatocellular carcinoma (HCC).

Methods: Expression of GNAI1 in HCC samples was determined by qRT-PCR and immunohistochemical (IHC) staining. Huh-7 and SNU-387 cells stably expressing GNAI1 were established by the infection of lentivirus transducing unit containing GNAI1. siRNA against GNAI1 was transfected into SMMC-7721 cells to knock down the GNAI1 expression in HCC cells. Mir-320a/c/d mimics were transfected into SMMC-7721 and SK-Hep-1 cells and the expression of GNAI1 was determined by Western blot. The migration and invasion of Huh-7, SNU-387, SK-Hep-1 and SMMC-7721 cells were investigated by Transwell assays.

Results: The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels. GNAI1 could inhibit the migration and invasion of HCC cells in vitro. Further investigations indicated that GNAI1 was a target of miR-320a/c/d in HCC cells. Transwell assays demonstrated that these microRNAs could promote the migratory ability and invasivesess of HCC cells in vitro.

Conclusions: GNAI1 is downregulated in HCC and inhibits the migration and invasion of HCC cells. This study is the first to investigate the role of GNAI1 in cancer. Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.

No MeSH data available.


Related in: MedlinePlus

GNAI1 is frequently down-regulated in HCC at the protein level but not at the mRNA level. A: Relative expression levels of GNAI1 in HCC and normal liver tissues using qRT-PCR assays. The box-plot lines represent the medians and interquartile ranges of the normalized threshold values (Ct), and the whiskers and spots indicate the 1st to 90th percentiles and the rest of the data points; B: Relative expression levels of GNAI1 in 50 HCC cases and matched noncancerous liver tissues; C: IHC staining of GNAI1 in HCC and matched noncancerous liver tissues (Non.) or in normal liver tissues (Liver). The staining intensities were scored and are represented as follows: a) 3; b) 0; c) 4; d) 1; e) 4; and f) 4. Statistical analysis compared groups of 10 cases of normal liver tissues and 50 cases of HCC to groups of matched noncancerous liver tissues according to the scoring results.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3643671&req=5

f1: GNAI1 is frequently down-regulated in HCC at the protein level but not at the mRNA level. A: Relative expression levels of GNAI1 in HCC and normal liver tissues using qRT-PCR assays. The box-plot lines represent the medians and interquartile ranges of the normalized threshold values (Ct), and the whiskers and spots indicate the 1st to 90th percentiles and the rest of the data points; B: Relative expression levels of GNAI1 in 50 HCC cases and matched noncancerous liver tissues; C: IHC staining of GNAI1 in HCC and matched noncancerous liver tissues (Non.) or in normal liver tissues (Liver). The staining intensities were scored and are represented as follows: a) 3; b) 0; c) 4; d) 1; e) 4; and f) 4. Statistical analysis compared groups of 10 cases of normal liver tissues and 50 cases of HCC to groups of matched noncancerous liver tissues according to the scoring results.

Mentions: To investigate whether the expression pattern of GNAI1 in HCC is similar to that of GNAI2, we measured both the mRNA and protein levels of GNAI1 in HCC samples and paired, adjacent noncancerous liver tissue samples from 50 patients as well as in normal liver tissue samples from 10 healthy patients. Although the mRNA level of GNAI1 was significantly lower in HCC than that in the normal liver samples (Figure 1A, P=0.0214), the GNAI1 mRNA levels showed no difference between the HCC and paired adjacent noncancerous liver tissues (Figure 1A, P=0.8372). However, an analysis of the GNAI1 protein expression in the HCC and normal liver tissues using IHC staining revealed that GNAI1 was significantly down-regulated in the HCC samples compared with the adjacent noncancerous liver tissues or the normal liver tissues (Figure 1B, HCC: n=50; normal liver: n=10). About two thirds of the HCC tissues (64%) had no or little expression of the GNAI1 protein. By contrast, the GNAI1 protein level was relatively high in all noncancerous and normal liver tissues (Figure 1C). The inconsistency between the GNAI1 expression at the mRNA and protein levels indicated the existence of post-transcriptional regulation, such as miRNA regulation. The GNAI1 expression in various HCC cell lines was also measured.


GNAI1 Suppresses Tumor Cell Migration and Invasion and is Post-Transcriptionally Regulated by Mir-320a/c/d in Hepatocellular Carcinoma.

Yao J, Liang LH, Zhang Y, Ding J, Tian Q, Li JJ, He XH - Cancer Biol Med (2012)

GNAI1 is frequently down-regulated in HCC at the protein level but not at the mRNA level. A: Relative expression levels of GNAI1 in HCC and normal liver tissues using qRT-PCR assays. The box-plot lines represent the medians and interquartile ranges of the normalized threshold values (Ct), and the whiskers and spots indicate the 1st to 90th percentiles and the rest of the data points; B: Relative expression levels of GNAI1 in 50 HCC cases and matched noncancerous liver tissues; C: IHC staining of GNAI1 in HCC and matched noncancerous liver tissues (Non.) or in normal liver tissues (Liver). The staining intensities were scored and are represented as follows: a) 3; b) 0; c) 4; d) 1; e) 4; and f) 4. Statistical analysis compared groups of 10 cases of normal liver tissues and 50 cases of HCC to groups of matched noncancerous liver tissues according to the scoring results.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3643671&req=5

f1: GNAI1 is frequently down-regulated in HCC at the protein level but not at the mRNA level. A: Relative expression levels of GNAI1 in HCC and normal liver tissues using qRT-PCR assays. The box-plot lines represent the medians and interquartile ranges of the normalized threshold values (Ct), and the whiskers and spots indicate the 1st to 90th percentiles and the rest of the data points; B: Relative expression levels of GNAI1 in 50 HCC cases and matched noncancerous liver tissues; C: IHC staining of GNAI1 in HCC and matched noncancerous liver tissues (Non.) or in normal liver tissues (Liver). The staining intensities were scored and are represented as follows: a) 3; b) 0; c) 4; d) 1; e) 4; and f) 4. Statistical analysis compared groups of 10 cases of normal liver tissues and 50 cases of HCC to groups of matched noncancerous liver tissues according to the scoring results.
Mentions: To investigate whether the expression pattern of GNAI1 in HCC is similar to that of GNAI2, we measured both the mRNA and protein levels of GNAI1 in HCC samples and paired, adjacent noncancerous liver tissue samples from 50 patients as well as in normal liver tissue samples from 10 healthy patients. Although the mRNA level of GNAI1 was significantly lower in HCC than that in the normal liver samples (Figure 1A, P=0.0214), the GNAI1 mRNA levels showed no difference between the HCC and paired adjacent noncancerous liver tissues (Figure 1A, P=0.8372). However, an analysis of the GNAI1 protein expression in the HCC and normal liver tissues using IHC staining revealed that GNAI1 was significantly down-regulated in the HCC samples compared with the adjacent noncancerous liver tissues or the normal liver tissues (Figure 1B, HCC: n=50; normal liver: n=10). About two thirds of the HCC tissues (64%) had no or little expression of the GNAI1 protein. By contrast, the GNAI1 protein level was relatively high in all noncancerous and normal liver tissues (Figure 1C). The inconsistency between the GNAI1 expression at the mRNA and protein levels indicated the existence of post-transcriptional regulation, such as miRNA regulation. The GNAI1 expression in various HCC cell lines was also measured.

Bottom Line: The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels.GNAI1 is downregulated in HCC and inhibits the migration and invasion of HCC cells.Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China ; State Key Laboratory for Diagnosis and Treatment for Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.

ABSTRACT

Objective: To explore the role and regulation of guanine nucleotide-binding protein G(i), α-1 subunit (GNAI1) in hepatocellular carcinoma (HCC).

Methods: Expression of GNAI1 in HCC samples was determined by qRT-PCR and immunohistochemical (IHC) staining. Huh-7 and SNU-387 cells stably expressing GNAI1 were established by the infection of lentivirus transducing unit containing GNAI1. siRNA against GNAI1 was transfected into SMMC-7721 cells to knock down the GNAI1 expression in HCC cells. Mir-320a/c/d mimics were transfected into SMMC-7721 and SK-Hep-1 cells and the expression of GNAI1 was determined by Western blot. The migration and invasion of Huh-7, SNU-387, SK-Hep-1 and SMMC-7721 cells were investigated by Transwell assays.

Results: The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels. GNAI1 could inhibit the migration and invasion of HCC cells in vitro. Further investigations indicated that GNAI1 was a target of miR-320a/c/d in HCC cells. Transwell assays demonstrated that these microRNAs could promote the migratory ability and invasivesess of HCC cells in vitro.

Conclusions: GNAI1 is downregulated in HCC and inhibits the migration and invasion of HCC cells. This study is the first to investigate the role of GNAI1 in cancer. Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.

No MeSH data available.


Related in: MedlinePlus