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An Lnc RNA (GAS5)/SnoRNA-derived piRNA induces activation of TRAIL gene by site-specifically recruiting MLL/COMPASS-like complexes.

He X, Chen X, Zhang X, Duan X, Pan T, Hu Q, Zhang Y, Zhong F, Liu J, Zhang H, Luo J, Wu K, Peng G, Luo H, Zhang L, Li X, Zhang H - Nucleic Acids Res. (2015)

Bottom Line: However, its function in the mammalian somatic cells remains unknown.Interestingly, the PIWIL1/4 proteins, which bind with this piRNA, directly interact with WDR5, resulting in a site-specific recruitment of the hCOMPASS-like complexes containing at least MLL3 and UTX (KDM6A).We have indicated a novel pathway for piRNAs to specially activate gene expression.

View Article: PubMed Central - PubMed

Affiliation: Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong 510080, China Key Laboratory of Tropical Disease Control of Ministry of Education, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong 510080, China.

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Synergistic effect of pi-sno75 and doxorubicin on apoptosis and on the growth of breast carcinoma cell lines in vitro and in vivo. (A) MCF7 cells were transfected with pi-sno75 or piNC; 48 h later, apoptosis was detected by FACS with Annexin-V/PI staining. (B) MCF7 cells were transfected by pi-sno75 or piNC. After 36 h, doxorubicin (Dox) was added to the cells for 12 h, and apoptosis was analysed by FACS. (C) After DR4 plus DR5 or WDR5 knockdown, impacts of pi-sno75 on apoptosis in MCF7 cells incubated with Dox were analyzed by FACS with Annexin-V staining. (D and E) After tumor formation without drug treatments, Immunohistochemistry (IHC) analysis of TRAIL expression in the MCF7 (D) or MDA-MB-231 (E) xenografts with pi-sno75 overexpression or with control vector. anti-TRAIL antibody was used as the primary antibody. (F and G) Tumor weights of MCF7 xenografts (F) or MDA-MB-231 (G) xenografts with pi-sno75 overexpression or control vector. The paired t-test was used. Data represent mean ± SD (error bars). *, statistically significant, ≤0.05.
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Figure 6: Synergistic effect of pi-sno75 and doxorubicin on apoptosis and on the growth of breast carcinoma cell lines in vitro and in vivo. (A) MCF7 cells were transfected with pi-sno75 or piNC; 48 h later, apoptosis was detected by FACS with Annexin-V/PI staining. (B) MCF7 cells were transfected by pi-sno75 or piNC. After 36 h, doxorubicin (Dox) was added to the cells for 12 h, and apoptosis was analysed by FACS. (C) After DR4 plus DR5 or WDR5 knockdown, impacts of pi-sno75 on apoptosis in MCF7 cells incubated with Dox were analyzed by FACS with Annexin-V staining. (D and E) After tumor formation without drug treatments, Immunohistochemistry (IHC) analysis of TRAIL expression in the MCF7 (D) or MDA-MB-231 (E) xenografts with pi-sno75 overexpression or with control vector. anti-TRAIL antibody was used as the primary antibody. (F and G) Tumor weights of MCF7 xenografts (F) or MDA-MB-231 (G) xenografts with pi-sno75 overexpression or control vector. The paired t-test was used. Data represent mean ± SD (error bars). *, statistically significant, ≤0.05.

Mentions: TRAIL has been well characterized as a specific tumor cell killer and serves as an effective anti-tumor agent (29). The upregulation of TRAIL expression by pi-sno75 suggests that the pi-sno75 could be of anti-tumor activity. Compared with the control, there was no apparent apoptosis in MCF7 cells transfected with pi-sno75 by Annexin-V/PI staining detected with FACS (Figure 6A). The result was in consistence with the observation that most breast cancer cells were relatively resistant to TRAIL-induced apoptosis (45). To avoid the tolerance for apoptosis, we treated the pi-sno75-transfected cells with doxorubicin (45,46). In comparison with a control small RNA, a significant increase in apoptosis took place in MCF7 cells treated with pi-sno75 under the same stimulation of doxorubicin (Figure 6B). The activity of pi-sno75 inducing cell death was counteracted by DR4 and DR5 knockdown (Figure 6C, middle panel). A similar phenomenon was also observed by the WDR5 knockdown (Figure 6C, low panel), further supporting the involvement of WDR5 and h-COMPASS complex.


An Lnc RNA (GAS5)/SnoRNA-derived piRNA induces activation of TRAIL gene by site-specifically recruiting MLL/COMPASS-like complexes.

He X, Chen X, Zhang X, Duan X, Pan T, Hu Q, Zhang Y, Zhong F, Liu J, Zhang H, Luo J, Wu K, Peng G, Luo H, Zhang L, Li X, Zhang H - Nucleic Acids Res. (2015)

Synergistic effect of pi-sno75 and doxorubicin on apoptosis and on the growth of breast carcinoma cell lines in vitro and in vivo. (A) MCF7 cells were transfected with pi-sno75 or piNC; 48 h later, apoptosis was detected by FACS with Annexin-V/PI staining. (B) MCF7 cells were transfected by pi-sno75 or piNC. After 36 h, doxorubicin (Dox) was added to the cells for 12 h, and apoptosis was analysed by FACS. (C) After DR4 plus DR5 or WDR5 knockdown, impacts of pi-sno75 on apoptosis in MCF7 cells incubated with Dox were analyzed by FACS with Annexin-V staining. (D and E) After tumor formation without drug treatments, Immunohistochemistry (IHC) analysis of TRAIL expression in the MCF7 (D) or MDA-MB-231 (E) xenografts with pi-sno75 overexpression or with control vector. anti-TRAIL antibody was used as the primary antibody. (F and G) Tumor weights of MCF7 xenografts (F) or MDA-MB-231 (G) xenografts with pi-sno75 overexpression or control vector. The paired t-test was used. Data represent mean ± SD (error bars). *, statistically significant, ≤0.05.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4402533&req=5

Figure 6: Synergistic effect of pi-sno75 and doxorubicin on apoptosis and on the growth of breast carcinoma cell lines in vitro and in vivo. (A) MCF7 cells were transfected with pi-sno75 or piNC; 48 h later, apoptosis was detected by FACS with Annexin-V/PI staining. (B) MCF7 cells were transfected by pi-sno75 or piNC. After 36 h, doxorubicin (Dox) was added to the cells for 12 h, and apoptosis was analysed by FACS. (C) After DR4 plus DR5 or WDR5 knockdown, impacts of pi-sno75 on apoptosis in MCF7 cells incubated with Dox were analyzed by FACS with Annexin-V staining. (D and E) After tumor formation without drug treatments, Immunohistochemistry (IHC) analysis of TRAIL expression in the MCF7 (D) or MDA-MB-231 (E) xenografts with pi-sno75 overexpression or with control vector. anti-TRAIL antibody was used as the primary antibody. (F and G) Tumor weights of MCF7 xenografts (F) or MDA-MB-231 (G) xenografts with pi-sno75 overexpression or control vector. The paired t-test was used. Data represent mean ± SD (error bars). *, statistically significant, ≤0.05.
Mentions: TRAIL has been well characterized as a specific tumor cell killer and serves as an effective anti-tumor agent (29). The upregulation of TRAIL expression by pi-sno75 suggests that the pi-sno75 could be of anti-tumor activity. Compared with the control, there was no apparent apoptosis in MCF7 cells transfected with pi-sno75 by Annexin-V/PI staining detected with FACS (Figure 6A). The result was in consistence with the observation that most breast cancer cells were relatively resistant to TRAIL-induced apoptosis (45). To avoid the tolerance for apoptosis, we treated the pi-sno75-transfected cells with doxorubicin (45,46). In comparison with a control small RNA, a significant increase in apoptosis took place in MCF7 cells treated with pi-sno75 under the same stimulation of doxorubicin (Figure 6B). The activity of pi-sno75 inducing cell death was counteracted by DR4 and DR5 knockdown (Figure 6C, middle panel). A similar phenomenon was also observed by the WDR5 knockdown (Figure 6C, low panel), further supporting the involvement of WDR5 and h-COMPASS complex.

Bottom Line: However, its function in the mammalian somatic cells remains unknown.Interestingly, the PIWIL1/4 proteins, which bind with this piRNA, directly interact with WDR5, resulting in a site-specific recruitment of the hCOMPASS-like complexes containing at least MLL3 and UTX (KDM6A).We have indicated a novel pathway for piRNAs to specially activate gene expression.

View Article: PubMed Central - PubMed

Affiliation: Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong 510080, China Key Laboratory of Tropical Disease Control of Ministry of Education, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong 510080, China.

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Related in: MedlinePlus