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The microRNA-218~Survivin axis regulates migration, invasion, and lymph node metastasis in cervical cancer.

Kogo R, How C, Chaudary N, Bruce J, Shi W, Hill RP, Zahedi P, Yip KW, Liu FF - Oncotarget (2015)

Bottom Line: Lower miR-218 expression was associated significantly with worse overall survival (OS), disease-free survival (DFS), and pelvic/aortic lymph node recurrence.In vitro, miR-218 over-expression decreased clonogenicity, migration, and invasion.YM155, a small molecule survivin inhibitor, significantly suppressed tumor growth and lymph node metastasis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Ontario Cancer Institute, University Health Network (UHN), Toronto, Ontario, Canada.

ABSTRACT
Cervical cancer is the third most common cancer in women worldwide. In the present study, global microRNA profiling for 79 cervical cancer patient samples led to the identification of miR-218 down-regulation in cervical cancer tissues compared to normal cervical tissues. Lower miR-218 expression was associated significantly with worse overall survival (OS), disease-free survival (DFS), and pelvic/aortic lymph node recurrence. In vitro, miR-218 over-expression decreased clonogenicity, migration, and invasion. Survivin (BIRC5) was subsequently identified as an important cervical cancer target of miR-218 using in silico prediction, mRNA profiling, and quantitative real-time PCR (qRT-PCR). Concordant with miR-218 over-expression, survivin knockdown by siRNA decreased clonogenicity, migration, and invasion. YM155, a small molecule survivin inhibitor, significantly suppressed tumor growth and lymph node metastasis in vivo. Our findings demonstrate that the miR-218~survivin axis inhibits cervical cancer progression by regulating clonogenicity, migration, and invasion, and suggest that the inhibition of survivin could be a potential therapeutic strategy to improve outcome in this disease.

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miR-218 reduces survival, migration, and invasion in vitroA) Clonogenic assays were performed by seeding SiHa and ME-180 cells transfected with 10 nM each of pre-miR negative control (miR-NC) or pre-miR-218 (miR-218). At 48 hrs post-transfection, cells were re-seeded at low density in 6-well plates. After 10-14 days incubation, colonies were stained and counted. B) Representative image (left) and quantification bar graph (right) of migrated SiHa and ME-180 cells. Migration assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in trans-well chambers. After 48 hrs incubation, the number of migrated cells were stained and counted. C) Representative image (left) and quantification bar graph (right) of invaded SiHa and ME-180 cells. Invasion assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in Matrigel invasion chambers. After 48 hrs incubation, the number of invaded cells were stained and counted. A-C: Bar graphs represent mean ± SEM from triplicates. *P<0.05, miR-NC: pre-miR negative control; miR-218: pre-miR-218.
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Figure 2: miR-218 reduces survival, migration, and invasion in vitroA) Clonogenic assays were performed by seeding SiHa and ME-180 cells transfected with 10 nM each of pre-miR negative control (miR-NC) or pre-miR-218 (miR-218). At 48 hrs post-transfection, cells were re-seeded at low density in 6-well plates. After 10-14 days incubation, colonies were stained and counted. B) Representative image (left) and quantification bar graph (right) of migrated SiHa and ME-180 cells. Migration assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in trans-well chambers. After 48 hrs incubation, the number of migrated cells were stained and counted. C) Representative image (left) and quantification bar graph (right) of invaded SiHa and ME-180 cells. Invasion assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in Matrigel invasion chambers. After 48 hrs incubation, the number of invaded cells were stained and counted. A-C: Bar graphs represent mean ± SEM from triplicates. *P<0.05, miR-NC: pre-miR negative control; miR-218: pre-miR-218.

Mentions: In order to elucidate the biological significance of miR-218 down-regulation, SiHa and ME-180 cells, which are both human papillomavirus (HPV) positive cervical squamous cell carcinoma lines, were transfected with pre-miR negative control (miR-NC) or pre-miR-218 (miR-218). Forty-eight hours post-transfection, miR-218 was over-expressed by more than 200-fold in SiHa and ME-180 cells (P<0.01, Figure S2). In both cell lines, this over-expression significantly reduced clonogenicity (P<0.05, Figure 2A).


The microRNA-218~Survivin axis regulates migration, invasion, and lymph node metastasis in cervical cancer.

Kogo R, How C, Chaudary N, Bruce J, Shi W, Hill RP, Zahedi P, Yip KW, Liu FF - Oncotarget (2015)

miR-218 reduces survival, migration, and invasion in vitroA) Clonogenic assays were performed by seeding SiHa and ME-180 cells transfected with 10 nM each of pre-miR negative control (miR-NC) or pre-miR-218 (miR-218). At 48 hrs post-transfection, cells were re-seeded at low density in 6-well plates. After 10-14 days incubation, colonies were stained and counted. B) Representative image (left) and quantification bar graph (right) of migrated SiHa and ME-180 cells. Migration assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in trans-well chambers. After 48 hrs incubation, the number of migrated cells were stained and counted. C) Representative image (left) and quantification bar graph (right) of invaded SiHa and ME-180 cells. Invasion assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in Matrigel invasion chambers. After 48 hrs incubation, the number of invaded cells were stained and counted. A-C: Bar graphs represent mean ± SEM from triplicates. *P<0.05, miR-NC: pre-miR negative control; miR-218: pre-miR-218.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 2: miR-218 reduces survival, migration, and invasion in vitroA) Clonogenic assays were performed by seeding SiHa and ME-180 cells transfected with 10 nM each of pre-miR negative control (miR-NC) or pre-miR-218 (miR-218). At 48 hrs post-transfection, cells were re-seeded at low density in 6-well plates. After 10-14 days incubation, colonies were stained and counted. B) Representative image (left) and quantification bar graph (right) of migrated SiHa and ME-180 cells. Migration assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in trans-well chambers. After 48 hrs incubation, the number of migrated cells were stained and counted. C) Representative image (left) and quantification bar graph (right) of invaded SiHa and ME-180 cells. Invasion assays were performed by seeding SiHa and ME-180 cells transfected with miR-NC or miR-218 (10 nM each) in Matrigel invasion chambers. After 48 hrs incubation, the number of invaded cells were stained and counted. A-C: Bar graphs represent mean ± SEM from triplicates. *P<0.05, miR-NC: pre-miR negative control; miR-218: pre-miR-218.
Mentions: In order to elucidate the biological significance of miR-218 down-regulation, SiHa and ME-180 cells, which are both human papillomavirus (HPV) positive cervical squamous cell carcinoma lines, were transfected with pre-miR negative control (miR-NC) or pre-miR-218 (miR-218). Forty-eight hours post-transfection, miR-218 was over-expressed by more than 200-fold in SiHa and ME-180 cells (P<0.01, Figure S2). In both cell lines, this over-expression significantly reduced clonogenicity (P<0.05, Figure 2A).

Bottom Line: Lower miR-218 expression was associated significantly with worse overall survival (OS), disease-free survival (DFS), and pelvic/aortic lymph node recurrence.In vitro, miR-218 over-expression decreased clonogenicity, migration, and invasion.YM155, a small molecule survivin inhibitor, significantly suppressed tumor growth and lymph node metastasis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Ontario Cancer Institute, University Health Network (UHN), Toronto, Ontario, Canada.

ABSTRACT
Cervical cancer is the third most common cancer in women worldwide. In the present study, global microRNA profiling for 79 cervical cancer patient samples led to the identification of miR-218 down-regulation in cervical cancer tissues compared to normal cervical tissues. Lower miR-218 expression was associated significantly with worse overall survival (OS), disease-free survival (DFS), and pelvic/aortic lymph node recurrence. In vitro, miR-218 over-expression decreased clonogenicity, migration, and invasion. Survivin (BIRC5) was subsequently identified as an important cervical cancer target of miR-218 using in silico prediction, mRNA profiling, and quantitative real-time PCR (qRT-PCR). Concordant with miR-218 over-expression, survivin knockdown by siRNA decreased clonogenicity, migration, and invasion. YM155, a small molecule survivin inhibitor, significantly suppressed tumor growth and lymph node metastasis in vivo. Our findings demonstrate that the miR-218~survivin axis inhibits cervical cancer progression by regulating clonogenicity, migration, and invasion, and suggest that the inhibition of survivin could be a potential therapeutic strategy to improve outcome in this disease.

Show MeSH
Related in: MedlinePlus