sh-apoc1  (Genechem)

Journal: Experimental and Therapeutic Medicine

Article Title: APOC1 promotes the progression of osteosarcoma by binding to MTCH2

doi: 10.3892/etm.2023.11862

Figure Lengend Snippet: APOC1 expression is significantly elevated in osteosarcoma cells and APOC1 silencing inhibits proliferation of osteosarcoma cells. Evaluation of APOC1 expression in osteosarcoma (HOS, SAOS-2, 143B and U2OS) and human osteoblast cell line hFOB1.19 using (A) RT-qPCR and (B) western blotting. *** P<0.001 vs. hFOB1.19. Assessment of APOC1 expression in SAOS-2 cells after transfection with sh-APOC1-1 or sh-APOC1-2 by (C) RT-qPCR and (D) western blotting. (E) Cell viability was tested by Cell Counting Kit-8 assay. (F) Proliferation of SAOS-2 cells was examined by colony formation assay. Magnification, x10. * P<0.05, *** P<0.001 vs. sh-NC. APOC1, apolipoprotein C1; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin; NC, negative control.

Article Snippet: For transfection, short hairpin RNAs (shRNAs) targeting APOC1 (sh-APOC1-1, 5'-TTCAGAAAGTGAAGGAGAAAC-3' and sh-APOC1-2, 5'-GTCTCCAGTGCCTTGGATAAG-3'), corresponding negative control (sh-NC, 5'-UUCUCCGAACGUGUCACGUTT-3'), MTCH2 pcDNA3.1 plasmid (Oe-MTCH2; cat. no. NM_001317231.2) and the empty vector control (Oe-NC) were supplied by Guangzhou RiboBio Co., Ltd. SAOS-2 cells were cultured in 6-well plates to 70% confluence at 37 ̊C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Cell Counting, Colony Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: APOC1 promotes the progression of osteosarcoma by binding to MTCH2

doi: 10.3892/etm.2023.11862

Figure Lengend Snippet: APOC1 silencing promotes apoptosis of osteosarcoma cells. (A) Cell apoptosis was detected using TUNEL staining. Magnification, x200. (B) Western blot analysis was to measure apoptosis-related protein expression. *** P<0.001 vs. sh-NC. APOC1, apolipoprotein C1; sh, short hairpin; NC, negative control.

Article Snippet: For transfection, short hairpin RNAs (shRNAs) targeting APOC1 (sh-APOC1-1, 5'-TTCAGAAAGTGAAGGAGAAAC-3' and sh-APOC1-2, 5'-GTCTCCAGTGCCTTGGATAAG-3'), corresponding negative control (sh-NC, 5'-UUCUCCGAACGUGUCACGUTT-3'), MTCH2 pcDNA3.1 plasmid (Oe-MTCH2; cat. no. NM_001317231.2) and the empty vector control (Oe-NC) were supplied by Guangzhou RiboBio Co., Ltd. SAOS-2 cells were cultured in 6-well plates to 70% confluence at 37 ̊C.

Techniques: TUNEL Assay, Staining, Western Blot, Expressing, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: APOC1 promotes the progression of osteosarcoma by binding to MTCH2

doi: 10.3892/etm.2023.11862

Figure Lengend Snippet: APOC1 silencing suppresses invasion and migration of osteosarcoma cells. (A) Invasion of SAOS-2 cells was determined by Transwell assay. Magnification, x100. (B) Wound healing assay was used to estimate cell migration. Magnification, x100. (C) Expression of MMP2 and MMP9 was detected by western blotting. ** P<0.01, *** P<0.001 vs. sh-NC. APOC1, apolipoprotein C1; MMP, matrix metallopeptidase; sh, short hairpin; NC, negative control.

Article Snippet: For transfection, short hairpin RNAs (shRNAs) targeting APOC1 (sh-APOC1-1, 5'-TTCAGAAAGTGAAGGAGAAAC-3' and sh-APOC1-2, 5'-GTCTCCAGTGCCTTGGATAAG-3'), corresponding negative control (sh-NC, 5'-UUCUCCGAACGUGUCACGUTT-3'), MTCH2 pcDNA3.1 plasmid (Oe-MTCH2; cat. no. NM_001317231.2) and the empty vector control (Oe-NC) were supplied by Guangzhou RiboBio Co., Ltd. SAOS-2 cells were cultured in 6-well plates to 70% confluence at 37 ̊C.

Techniques: Migration, Transwell Assay, Wound Healing Assay, Expressing, Western Blot, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: APOC1 promotes the progression of osteosarcoma by binding to MTCH2

doi: 10.3892/etm.2023.11862

Figure Lengend Snippet: APOC1 may interact with MTCH2 in osteosarcoma. (A) BioGRID and (B) STRING databases predicted that APOC1 interacted with MTCH2. Measurement of MTCH2 expression in SAOS-2 and hFOB1.19 cells by (C) reverse transcription-quantitative PCR and (D) western blotting. *** P<0.001 vs. hFOB1.19. (E) Verification of the interaction between APOC1 and MTCH2 using co-immunoprecipitation assay. APOC1, apolipoprotein C1; MTCH2, mitochondrial carrier homolog 2.

Article Snippet: For transfection, short hairpin RNAs (shRNAs) targeting APOC1 (sh-APOC1-1, 5'-TTCAGAAAGTGAAGGAGAAAC-3' and sh-APOC1-2, 5'-GTCTCCAGTGCCTTGGATAAG-3'), corresponding negative control (sh-NC, 5'-UUCUCCGAACGUGUCACGUTT-3'), MTCH2 pcDNA3.1 plasmid (Oe-MTCH2; cat. no. NM_001317231.2) and the empty vector control (Oe-NC) were supplied by Guangzhou RiboBio Co., Ltd. SAOS-2 cells were cultured in 6-well plates to 70% confluence at 37 ̊C.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Co-Immunoprecipitation Assay

Journal: Experimental and Therapeutic Medicine

Article Title: APOC1 promotes the progression of osteosarcoma by binding to MTCH2

doi: 10.3892/etm.2023.11862

Figure Lengend Snippet: MTCH2 overexpression inhibits the effect of APOC1 deletion on the proliferation and apoptosis of osteosarcoma cells. Expression of MTCH2 after transfection with Oe-MTCH2 was detected using (A) reverse transcription-quantitative PCR and (B) western blotting. *** P<0.001 vs. Oe-NC. (C) Cell viability was analyzed by Cell Counting Kit-8 assay. (D) Colony formation assay was used for the detection of cell proliferation. Magnification, x10. (E) Cell apoptosis was tested using TUNEL staining. Magnification, x200. (F) Expression of apoptosis-related proteins was estimated using western blotting. *** P<0.001 vs. sh-NC; # P<0.05, ## P<0.01, ### P<0.001 vs. sh-APOC1 + Oe-NC. MTCH2, mitochondrial carrier homolog 2; APOC1, apolipoprotein C1; Oe, overexpression; NC, negative control; sh, short hairpin.

Article Snippet: For transfection, short hairpin RNAs (shRNAs) targeting APOC1 (sh-APOC1-1, 5'-TTCAGAAAGTGAAGGAGAAAC-3' and sh-APOC1-2, 5'-GTCTCCAGTGCCTTGGATAAG-3'), corresponding negative control (sh-NC, 5'-UUCUCCGAACGUGUCACGUTT-3'), MTCH2 pcDNA3.1 plasmid (Oe-MTCH2; cat. no. NM_001317231.2) and the empty vector control (Oe-NC) were supplied by Guangzhou RiboBio Co., Ltd. SAOS-2 cells were cultured in 6-well plates to 70% confluence at 37 ̊C.

Techniques: Over Expression, Expressing, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Cell Counting, Colony Assay, TUNEL Assay, Staining, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: APOC1 promotes the progression of osteosarcoma by binding to MTCH2

doi: 10.3892/etm.2023.11862

Figure Lengend Snippet: MTCH2 overexpression attenuates the effects of APOC1 knockdown on invasion and migration of osteosarcoma cells. (A) Invasion of SAOS-2 cells was evaluated by Transwell assay. Magnification, x100. (B) Wound healing assay was used for measurement of cell migration. Magnification, x100. (C) Expression of MMP2 and MMP9 was determined using western blotting. *** P<0.001 vs. sh-NC; # P<0.05, ## P<0.01, ### P<0.001 vs. sh-APOC1 + Oe-NC. MTCH2, mitochondrial carrier homolog 2; APOC1, apolipoprotein C1; MMP, matrix metallopeptidase; sh, short hairpin; NC, negative control; Oe, overexpression.

Article Snippet: For transfection, short hairpin RNAs (shRNAs) targeting APOC1 (sh-APOC1-1, 5'-TTCAGAAAGTGAAGGAGAAAC-3' and sh-APOC1-2, 5'-GTCTCCAGTGCCTTGGATAAG-3'), corresponding negative control (sh-NC, 5'-UUCUCCGAACGUGUCACGUTT-3'), MTCH2 pcDNA3.1 plasmid (Oe-MTCH2; cat. no. NM_001317231.2) and the empty vector control (Oe-NC) were supplied by Guangzhou RiboBio Co., Ltd. SAOS-2 cells were cultured in 6-well plates to 70% confluence at 37 ̊C.

Techniques: Over Expression, Knockdown, Migration, Transwell Assay, Wound Healing Assay, Expressing, Western Blot, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: APOC1 promotes the progression of osteosarcoma by binding to MTCH2

doi: 10.3892/etm.2023.11862

Figure Lengend Snippet: APOC1 silencing promotes oxidative phosphorylation and inhibits the Warburg effect of osteosarcoma cells by regulating MTCH2 expression. (A) OCR of SAOS-2 cells was detected using the Seahorse XF Cell Mito Stress Test kit. (B) ECAR was assessed using the Seahorse XF Glycolysis Stress Test kit. (C) Lactate production was tested using lactate assay kit. Corresponding commercially available kits were used to measure the activity of (D) HK, (E) PFK and (F) PK. * P<0.05, ** P<0.01, *** P<0.001 vs. sh-NC; # P<0.05, ## P<0.01, ### P<0.001 vs. sh-APOC1 + Oe-NC. APOC1, apolipoprotein C1; MTCH2, mitochondrial carrier homolog 2; OCR, O 2 consumption rate; ECAR, extracellular acidification rate; HK, hexokinase; PFK, phosphofructokinase; PK, pyruvate kinase; sh, short hairpin; NC, negative control; Oe, overexpression.

Article Snippet: For transfection, short hairpin RNAs (shRNAs) targeting APOC1 (sh-APOC1-1, 5'-TTCAGAAAGTGAAGGAGAAAC-3' and sh-APOC1-2, 5'-GTCTCCAGTGCCTTGGATAAG-3'), corresponding negative control (sh-NC, 5'-UUCUCCGAACGUGUCACGUTT-3'), MTCH2 pcDNA3.1 plasmid (Oe-MTCH2; cat. no. NM_001317231.2) and the empty vector control (Oe-NC) were supplied by Guangzhou RiboBio Co., Ltd. SAOS-2 cells were cultured in 6-well plates to 70% confluence at 37 ̊C.

Techniques: Phospho-proteomics, Expressing, Lactate Assay, Activity Assay, Negative Control, Over Expression