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REV3L, a promising target in regulating the chemosensitivity of cervical cancer cells.

Yang L, Shi T, Liu F, Ren C, Wang Z, Li Y, Tu X, Yang G, Cheng X - PLoS ONE (2015)

Bottom Line: REV3L, the catalytic subunit of DNA Polymerase ζ (Polζ), plays a significant role in the DNA damage tolerance mechanism of translesion synthesis (TLS).The suppression of REV3L expression enhanced the sensitivity of cervical cancer cells to cisplatin, and the overexpression of REV3L conferred resistance to cisplatin as evidenced by the alteration of apoptosis rates, and significantly expression level changes of anti-apoptotic proteins B-cell lymphoma 2 (Bcl-2), myeloid cell leukemia sequence 1 (Mcl-1) and B-cell lymphoma-extra large (Bcl-xl) and proapoptotic Bcl-2-associated x protein (Bax).Our data suggest that REV3L plays an important role in regulating cervical cancer cellular response to cisplatin, and thus targeting REV3L may be a promising way to alter chemosensitivity in cervical cancer patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Gynecologic Oncology, Zhejiang Cancer Hospital, Hangzhou, Zhejiang, China.

ABSTRACT
REV3L, the catalytic subunit of DNA Polymerase ζ (Polζ), plays a significant role in the DNA damage tolerance mechanism of translesion synthesis (TLS). The role of REV3L in chemosensitivity of cervical cancer needs exploration. In the present study, we evaluated the expression of the Polζ protein in paraffin-embedded tissues using immunohistochemistry and found that the expression of Polζ in cervical cancer tissues was higher than that in normal tissues. We then established some cervical cancer cell lines with REV3L suppression or overexpression. Depletion of REV3L suppresses cell proliferation and colony formation of cervical cancer cells through G1 arrest, and REV3L promotes cell proliferation and colony formation of cervical cancer cells by promoting G1 phase to S phase transition. The suppression of REV3L expression enhanced the sensitivity of cervical cancer cells to cisplatin, and the overexpression of REV3L conferred resistance to cisplatin as evidenced by the alteration of apoptosis rates, and significantly expression level changes of anti-apoptotic proteins B-cell lymphoma 2 (Bcl-2), myeloid cell leukemia sequence 1 (Mcl-1) and B-cell lymphoma-extra large (Bcl-xl) and proapoptotic Bcl-2-associated x protein (Bax). Our data suggest that REV3L plays an important role in regulating cervical cancer cellular response to cisplatin, and thus targeting REV3L may be a promising way to alter chemosensitivity in cervical cancer patients.

No MeSH data available.


Related in: MedlinePlus

Alteration of mitochondria-associated apoptotic proteins after cisplatin treatment.(A)The expression levels of Bcl-2, Bcl-xl and Mcl-1 were lower and levels of Bax and cytochrome c were higher in shREV3L cells compared with shGFP cells in response to the same dose of cisplatin. REV3L-overexpressing MS751 and ME180 cells showed higher levels of Bcl-2, Mcl-1 and Bcl-xl and lower levels of Bax and cytochrome c compared to the vector control cells. (B) γ-H2AX and p-p53(pS15) proteins were increased in SiHa shREV3L cells and decreased in ME180 REV3L cells, compared with control cells after cisplatin treatment. (C) Time-dependent expression of cleaved caspase-3 after exposure to a single dose of cisplatin (1 μmol/L) in MS751 cells and a single dose of cisplatin (10 μmol/L) in HeLa cells. Expression levels of cleaved caspase-3 were lower in MS751 REV3L cells and higher in HeLa shREV3L cells compared with control cells in response to the same dose of cisplatin in a time-dependent manner. (D) Normalized fold expression of each protein against internal control protein. Data are means of three independent experiments ± SEM. * P< 0.05.
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pone.0120334.g005: Alteration of mitochondria-associated apoptotic proteins after cisplatin treatment.(A)The expression levels of Bcl-2, Bcl-xl and Mcl-1 were lower and levels of Bax and cytochrome c were higher in shREV3L cells compared with shGFP cells in response to the same dose of cisplatin. REV3L-overexpressing MS751 and ME180 cells showed higher levels of Bcl-2, Mcl-1 and Bcl-xl and lower levels of Bax and cytochrome c compared to the vector control cells. (B) γ-H2AX and p-p53(pS15) proteins were increased in SiHa shREV3L cells and decreased in ME180 REV3L cells, compared with control cells after cisplatin treatment. (C) Time-dependent expression of cleaved caspase-3 after exposure to a single dose of cisplatin (1 μmol/L) in MS751 cells and a single dose of cisplatin (10 μmol/L) in HeLa cells. Expression levels of cleaved caspase-3 were lower in MS751 REV3L cells and higher in HeLa shREV3L cells compared with control cells in response to the same dose of cisplatin in a time-dependent manner. (D) Normalized fold expression of each protein against internal control protein. Data are means of three independent experiments ± SEM. * P< 0.05.

Mentions: As the mitochondria-associated apoptotic pathway participates in cellular response in tumors to many anticancer drugs, to see whether it is involved in REV3L induced alteration in sensitivity to cisplatin, we performed an analysis of proapoptotic proteins (Bax, cytochrome c) and antiapoptotic proteins (Bcl-2, Bcl-xl, and Mcl-1) in the vector control cells and the REV3L-overexpressing or suppression cells with cisplatin treatment (Fig. 5A, D). After treatment with cisplatin for 24 h, Bcl-2, Bcl-xl and Mcl-1 levels markedly decreased, and Bax and cytochrome c levels increased in shREV3L HeLa and SiHa cells, compared with shGFP HeLa and SiHa cells. Consistently, after treatment with cisplatin, REV3L-overexpressing MS751 and ME180 cells showed higher levels of Bcl-2, Mcl-1 and Bcl-xl and lower levels of Bax and cytochrome c, compared to the vector control cells. Furthermore, after exposure to cisplatin (1 μmol/L) for 72 h, a time dependent increase in the amounts of cleaved caspase-3 was observed in HeLa shREV3L cells and a decrease in the amounts of cleaved caspase-3 was observed in MS751 REV3L cells after a single dose treatment of 10 μmol/L of cisplatin (Fig. 5C, D). Thus, these results suggest that REV3L confers chemoresistance to cisplatin through the mitochondria- associated apoptotic pathway.


REV3L, a promising target in regulating the chemosensitivity of cervical cancer cells.

Yang L, Shi T, Liu F, Ren C, Wang Z, Li Y, Tu X, Yang G, Cheng X - PLoS ONE (2015)

Alteration of mitochondria-associated apoptotic proteins after cisplatin treatment.(A)The expression levels of Bcl-2, Bcl-xl and Mcl-1 were lower and levels of Bax and cytochrome c were higher in shREV3L cells compared with shGFP cells in response to the same dose of cisplatin. REV3L-overexpressing MS751 and ME180 cells showed higher levels of Bcl-2, Mcl-1 and Bcl-xl and lower levels of Bax and cytochrome c compared to the vector control cells. (B) γ-H2AX and p-p53(pS15) proteins were increased in SiHa shREV3L cells and decreased in ME180 REV3L cells, compared with control cells after cisplatin treatment. (C) Time-dependent expression of cleaved caspase-3 after exposure to a single dose of cisplatin (1 μmol/L) in MS751 cells and a single dose of cisplatin (10 μmol/L) in HeLa cells. Expression levels of cleaved caspase-3 were lower in MS751 REV3L cells and higher in HeLa shREV3L cells compared with control cells in response to the same dose of cisplatin in a time-dependent manner. (D) Normalized fold expression of each protein against internal control protein. Data are means of three independent experiments ± SEM. * P< 0.05.
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pone.0120334.g005: Alteration of mitochondria-associated apoptotic proteins after cisplatin treatment.(A)The expression levels of Bcl-2, Bcl-xl and Mcl-1 were lower and levels of Bax and cytochrome c were higher in shREV3L cells compared with shGFP cells in response to the same dose of cisplatin. REV3L-overexpressing MS751 and ME180 cells showed higher levels of Bcl-2, Mcl-1 and Bcl-xl and lower levels of Bax and cytochrome c compared to the vector control cells. (B) γ-H2AX and p-p53(pS15) proteins were increased in SiHa shREV3L cells and decreased in ME180 REV3L cells, compared with control cells after cisplatin treatment. (C) Time-dependent expression of cleaved caspase-3 after exposure to a single dose of cisplatin (1 μmol/L) in MS751 cells and a single dose of cisplatin (10 μmol/L) in HeLa cells. Expression levels of cleaved caspase-3 were lower in MS751 REV3L cells and higher in HeLa shREV3L cells compared with control cells in response to the same dose of cisplatin in a time-dependent manner. (D) Normalized fold expression of each protein against internal control protein. Data are means of three independent experiments ± SEM. * P< 0.05.
Mentions: As the mitochondria-associated apoptotic pathway participates in cellular response in tumors to many anticancer drugs, to see whether it is involved in REV3L induced alteration in sensitivity to cisplatin, we performed an analysis of proapoptotic proteins (Bax, cytochrome c) and antiapoptotic proteins (Bcl-2, Bcl-xl, and Mcl-1) in the vector control cells and the REV3L-overexpressing or suppression cells with cisplatin treatment (Fig. 5A, D). After treatment with cisplatin for 24 h, Bcl-2, Bcl-xl and Mcl-1 levels markedly decreased, and Bax and cytochrome c levels increased in shREV3L HeLa and SiHa cells, compared with shGFP HeLa and SiHa cells. Consistently, after treatment with cisplatin, REV3L-overexpressing MS751 and ME180 cells showed higher levels of Bcl-2, Mcl-1 and Bcl-xl and lower levels of Bax and cytochrome c, compared to the vector control cells. Furthermore, after exposure to cisplatin (1 μmol/L) for 72 h, a time dependent increase in the amounts of cleaved caspase-3 was observed in HeLa shREV3L cells and a decrease in the amounts of cleaved caspase-3 was observed in MS751 REV3L cells after a single dose treatment of 10 μmol/L of cisplatin (Fig. 5C, D). Thus, these results suggest that REV3L confers chemoresistance to cisplatin through the mitochondria- associated apoptotic pathway.

Bottom Line: REV3L, the catalytic subunit of DNA Polymerase ζ (Polζ), plays a significant role in the DNA damage tolerance mechanism of translesion synthesis (TLS).The suppression of REV3L expression enhanced the sensitivity of cervical cancer cells to cisplatin, and the overexpression of REV3L conferred resistance to cisplatin as evidenced by the alteration of apoptosis rates, and significantly expression level changes of anti-apoptotic proteins B-cell lymphoma 2 (Bcl-2), myeloid cell leukemia sequence 1 (Mcl-1) and B-cell lymphoma-extra large (Bcl-xl) and proapoptotic Bcl-2-associated x protein (Bax).Our data suggest that REV3L plays an important role in regulating cervical cancer cellular response to cisplatin, and thus targeting REV3L may be a promising way to alter chemosensitivity in cervical cancer patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Gynecologic Oncology, Zhejiang Cancer Hospital, Hangzhou, Zhejiang, China.

ABSTRACT
REV3L, the catalytic subunit of DNA Polymerase ζ (Polζ), plays a significant role in the DNA damage tolerance mechanism of translesion synthesis (TLS). The role of REV3L in chemosensitivity of cervical cancer needs exploration. In the present study, we evaluated the expression of the Polζ protein in paraffin-embedded tissues using immunohistochemistry and found that the expression of Polζ in cervical cancer tissues was higher than that in normal tissues. We then established some cervical cancer cell lines with REV3L suppression or overexpression. Depletion of REV3L suppresses cell proliferation and colony formation of cervical cancer cells through G1 arrest, and REV3L promotes cell proliferation and colony formation of cervical cancer cells by promoting G1 phase to S phase transition. The suppression of REV3L expression enhanced the sensitivity of cervical cancer cells to cisplatin, and the overexpression of REV3L conferred resistance to cisplatin as evidenced by the alteration of apoptosis rates, and significantly expression level changes of anti-apoptotic proteins B-cell lymphoma 2 (Bcl-2), myeloid cell leukemia sequence 1 (Mcl-1) and B-cell lymphoma-extra large (Bcl-xl) and proapoptotic Bcl-2-associated x protein (Bax). Our data suggest that REV3L plays an important role in regulating cervical cancer cellular response to cisplatin, and thus targeting REV3L may be a promising way to alter chemosensitivity in cervical cancer patients.

No MeSH data available.


Related in: MedlinePlus