Impact of Bupivacaine on malignant proliferation, apoptosis and autophagy of human colorectal cancer SW480 cells through regulating NF-κB signaling path

Figures & data

Table 1. qRT-PCR primer sequence

	Primer sequence (5’3’)
GAPDH	Forward: 5’-CCTCGTCTCATAGACAAGATGGT-3’
	Reserse: 5’-GGGTAGAGTCATACTGGAACATG-3’
Bcl-2	Forward: 5’- CTGGTGGACAACATCGCTCTG −3’
	Reserse: 5’- GGTCTGCTGACCTCACTTGTG −3’
Bax	Forward: 5’- GGATCGAGCAGAGAGGATGG −3’
	Reserse: 5’- TGGTGAGTGAGGCAGTGAGG −3’
Caspase-3	Forward: 5’- GGATCGAGCAGAGAGGATGG −3’
	Reserse: 5’- TGGTGAGTGAGGCAGTGAGG −3’

Figure 1. Bupivacaine Inhibited CRC Cell Proliferation

Note: A & B: MTT Assay For Checking SW480 Cell Activity After Receiving 1 μm-1 mm Bupivacaine In 24 And 48 h; C & D:MTT Assay For Checking SW620 Cell Activity After Receiving 1 μm-1 mm Bupivacaine In 24 And 48 h; E & F:MTT Assay For Checking FHC Cell Activity After Receiving 1 μm-1 mm Bupivacaine In 24 And 48 h; Data Shown In Mean ± SD (N = 3); One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05.

Figure 2. Bupivacaine Inhibited CRC Cell Migration

Note: A & B: Cell Scratch Test For Checking The Cell Migration Capacity In In SW480 And SW620 Cells After Receiving 1 mm Bupivacaine In 24 And 48 h; Data Shown In Mean ± SD (N = 3); One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05.

Figure 3. Bupivacaine Promoted CRC Cell Apoptosis

Note: A & B: Flow Cytometry For Checking The Cell Apoptosis Rete In In SW480 And SW620 Cells After Receiving 1 mm Bupivacaine In 24 And 48 h; C & D:Qrt-PCR For Checking Bcl-2, Bax And Caspase-3 Mrna Expression Levels In SW480 And SW620 Cells After Receiving 1 mm Bupivacaine In 24 h And 48 h; Data Shown In Mean ± SD (N = 3); One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05.

Figure 4. Bupivacaine Promoted CRC Cell Autophagy

Note: A: Western Blot For Checking LC3B, Beclin-1 And P62 Protein Expression Levels In SW480 And SW620 Cells After Receiving 1 mm Bupivacaine In 24 h; B: Western Blot For Checking LC3B, Beclin-1 And P62 Protein Expression Levels In SW480 And SW620 Cells After Receiving 1 mm Bupivacaine In 48 h; Data Shown In Mean ± SD (N = 3); One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05.

Figure 5. Bupivacaine Inhibited NF-Κb Activation In CRC Cells

Note: A & B: Immunofluorescence For Checking NF-Κb Activation In SW480 Cells After Receiving 1 mm Bupivacaine In 24 h Or 48 h; C & D: Immunofluorescence For Checking NF-Κb Activation In SW620 Cells After Receiving 1 mm Bupivacaine In 24 h And 48 h; Data Shown In Mean ±SD (N = 3); One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05.

Figure 6. Bupivacaine Inhibited CRC Inflammation Factors

Note: A & B: ELISA For Checking TNF-Α, IL-1β, And IL-6 Levels In SW480 And SW620 Cells After Receiving 1 mm Bupivacaine In 24 h And 48 h; One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05.

Figure 7. Bupivacaine Inhibited CRC NF-Κb Signaling Path

Note: A & B: Western Blot For Checking NF-Κb, Iκb Phosphorylation And IKK Phosphorylation Protein Expression In SW480 And SW620 Cells After Receiving 1 mm Bupivacaine; Data Shown In Mean ± SD (N = 3); One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05.

Figure 8. Bupivacaine Inhibited Tumor Growth In Body

Note: A: Photos, Volume And Weight Of Mouse Tumors After Receiving 4.3 mm And 8.6 mm Bupivacaine; B: Western Blot For Detecting NF-Κb Expression After Receiving 4.3 mm And 8.6 mm Bupivacaine; Data Shown In Mean ± SD (N = 6). One-Way ANOVA For Calculating The Significance Of Each Group; Applying The Tukey Test For Correcting The Variance. *P< 0.05。