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HPV18 E7 induces the over-transcription of eIF4E gene in cervical cancer.

Pang T, Wang S, Gao M, Kang H, Zhao Y, Yao Y, Hu X - Iran J Basic Med Sci (2015)

Bottom Line: Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells.HPV E7 induced eIF4E gene over transcription which might be a new marker for CC.The finding broadens the understanding of the CC carcinogenesis.

View Article: PubMed Central - PubMed

Affiliation: Obstetrics and Gynecology Department of the first Affiliated Hospital of Guangdong Medical College, No. 57 Avenue of the people, Zhanjiang, Guangdong Province 524023, PR China ; Cancer Institute of Guangdong Medical College, No. 1 Xincheng Road, Dongguan, Guangdong Province 523808, PR China.

ABSTRACT

Objectives: Eukaryotic translation initiation factor 4E (eIF4E) is overexpressed in cervical cancer (CC). However, the molecular mechanisms are unclear. This study aimed to investigate the molecular mechanism of eIF4E gene overexpression in CC.

Materials and methods: The human papillomavirus (HPV) type 18 E7 and eIF4E mRNAs were measured following knock down or overexpression of E7 gene by RT-PCR and real-time PCR. Cell counting kit-8 assay was used to determine the cell proliferation. Flow cytometry was used to analyze the cell cycle and apoptosis. Transwell system was employed to determine the cell migration.

Results: Overexpression of E7 gene increased eIF4E mRNA level by 24.3% (P<0.01) in HPV negative C33A cells. Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells. Under the state of high expression of E7, 1) up-regulation of eIF4E drastically promoted the cell proliferation, cell cycle and cell migration, and inhibited the cell apoptosis. 2) down-regulation of eIF4E significantly inhibited the cell proliferation, cell cycle and the ability of cell migration, and also promoted the apoptosis of cervical cancer cells.

Conclusion: HPV E7 induced eIF4E gene over transcription which might be a new marker for CC. The finding broadens the understanding of the CC carcinogenesis.

No MeSH data available.


Related in: MedlinePlus

Knockdown of e7 in HeLa cells down regulated eukaryotic translation initiation factor 4e gene expression. (A) E7 mRNA expression was decreased by shE7s at 48 hr detected by real-time PCR. (B) Detection of E7 mRNA expression by RT-PCR; (C) eIF4E mRNA expression decreased after E7 knockdown at 48 hr detected by real-time PCR. (D) Detection of eIF4E mRNA expression by RT-PCR; (E) eIF4E protein expression decreased in HeLa cells at 24 hr and 48 hr after the transfection of shE7. Mock, HeLa cells; NC, blank vector group; *: vs Mock, P <0.05; **: vs Mock, P<0.01
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Figure 2: Knockdown of e7 in HeLa cells down regulated eukaryotic translation initiation factor 4e gene expression. (A) E7 mRNA expression was decreased by shE7s at 48 hr detected by real-time PCR. (B) Detection of E7 mRNA expression by RT-PCR; (C) eIF4E mRNA expression decreased after E7 knockdown at 48 hr detected by real-time PCR. (D) Detection of eIF4E mRNA expression by RT-PCR; (E) eIF4E protein expression decreased in HeLa cells at 24 hr and 48 hr after the transfection of shE7. Mock, HeLa cells; NC, blank vector group; *: vs Mock, P <0.05; **: vs Mock, P<0.01

Mentions: The shE7 vectors carrying GFP were successfully constructed. The E7 mRNA level in the NC group was similar with that in the mock group, determined by both real-time PCR and RT-PCR test. By the real-time PCR detection, the E7 mRNA in shE7-1, 2, 3 treated groups were significantly lower than that of the NC group (Figure 2A). Among shE7-1, 2, 3, shE7-2 showed the most effective inhibition of E7 mRNA, with an inhibition rate of approximately 81%. By the RT-PCR and agarose gel detection, two distinct bands with expected sizes were seen for E7 gene (119bp) and GAPDH gene (224 bp) in all groups (Figure 2B). By Image J band analysis, the E7 mRNA in shE7-1 and shE7-2 but not shE7-3 groups was considerably knocked down, compared with the NC group. The E7 mRNA level in shE7-2 group was lower than that in shE7-1 group (Figure 2B). The changes of the E7 mRNA were comparable in both the real-time PCR and RT-PCR detection (Figure 2).


HPV18 E7 induces the over-transcription of eIF4E gene in cervical cancer.

Pang T, Wang S, Gao M, Kang H, Zhao Y, Yao Y, Hu X - Iran J Basic Med Sci (2015)

Knockdown of e7 in HeLa cells down regulated eukaryotic translation initiation factor 4e gene expression. (A) E7 mRNA expression was decreased by shE7s at 48 hr detected by real-time PCR. (B) Detection of E7 mRNA expression by RT-PCR; (C) eIF4E mRNA expression decreased after E7 knockdown at 48 hr detected by real-time PCR. (D) Detection of eIF4E mRNA expression by RT-PCR; (E) eIF4E protein expression decreased in HeLa cells at 24 hr and 48 hr after the transfection of shE7. Mock, HeLa cells; NC, blank vector group; *: vs Mock, P <0.05; **: vs Mock, P<0.01
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4556762&req=5

Figure 2: Knockdown of e7 in HeLa cells down regulated eukaryotic translation initiation factor 4e gene expression. (A) E7 mRNA expression was decreased by shE7s at 48 hr detected by real-time PCR. (B) Detection of E7 mRNA expression by RT-PCR; (C) eIF4E mRNA expression decreased after E7 knockdown at 48 hr detected by real-time PCR. (D) Detection of eIF4E mRNA expression by RT-PCR; (E) eIF4E protein expression decreased in HeLa cells at 24 hr and 48 hr after the transfection of shE7. Mock, HeLa cells; NC, blank vector group; *: vs Mock, P <0.05; **: vs Mock, P<0.01
Mentions: The shE7 vectors carrying GFP were successfully constructed. The E7 mRNA level in the NC group was similar with that in the mock group, determined by both real-time PCR and RT-PCR test. By the real-time PCR detection, the E7 mRNA in shE7-1, 2, 3 treated groups were significantly lower than that of the NC group (Figure 2A). Among shE7-1, 2, 3, shE7-2 showed the most effective inhibition of E7 mRNA, with an inhibition rate of approximately 81%. By the RT-PCR and agarose gel detection, two distinct bands with expected sizes were seen for E7 gene (119bp) and GAPDH gene (224 bp) in all groups (Figure 2B). By Image J band analysis, the E7 mRNA in shE7-1 and shE7-2 but not shE7-3 groups was considerably knocked down, compared with the NC group. The E7 mRNA level in shE7-2 group was lower than that in shE7-1 group (Figure 2B). The changes of the E7 mRNA were comparable in both the real-time PCR and RT-PCR detection (Figure 2).

Bottom Line: Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells.HPV E7 induced eIF4E gene over transcription which might be a new marker for CC.The finding broadens the understanding of the CC carcinogenesis.

View Article: PubMed Central - PubMed

Affiliation: Obstetrics and Gynecology Department of the first Affiliated Hospital of Guangdong Medical College, No. 57 Avenue of the people, Zhanjiang, Guangdong Province 524023, PR China ; Cancer Institute of Guangdong Medical College, No. 1 Xincheng Road, Dongguan, Guangdong Province 523808, PR China.

ABSTRACT

Objectives: Eukaryotic translation initiation factor 4E (eIF4E) is overexpressed in cervical cancer (CC). However, the molecular mechanisms are unclear. This study aimed to investigate the molecular mechanism of eIF4E gene overexpression in CC.

Materials and methods: The human papillomavirus (HPV) type 18 E7 and eIF4E mRNAs were measured following knock down or overexpression of E7 gene by RT-PCR and real-time PCR. Cell counting kit-8 assay was used to determine the cell proliferation. Flow cytometry was used to analyze the cell cycle and apoptosis. Transwell system was employed to determine the cell migration.

Results: Overexpression of E7 gene increased eIF4E mRNA level by 24.3% (P<0.01) in HPV negative C33A cells. Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells. Under the state of high expression of E7, 1) up-regulation of eIF4E drastically promoted the cell proliferation, cell cycle and cell migration, and inhibited the cell apoptosis. 2) down-regulation of eIF4E significantly inhibited the cell proliferation, cell cycle and the ability of cell migration, and also promoted the apoptosis of cervical cancer cells.

Conclusion: HPV E7 induced eIF4E gene over transcription which might be a new marker for CC. The finding broadens the understanding of the CC carcinogenesis.

No MeSH data available.


Related in: MedlinePlus