5092c51d544159873492357c3a9fb0e2 ijms-25-12997.pdf f365bf6e8f5626f9fb2106d617e9feee13c972a2 ijms-25-12997.pdf 9e87a8b76b15725ca4e87e25dd4e5a3be1809f37e499c7494be04b963c01338f ijms-25-12997.pdf Title: Mycobacteria Treatment Inhibits Bladder Cancer Cell Migration, Invasion, and Anchorage-Independent Growth Subject: Bladder cancer (BC) is a highly recurrent and invasive malignancy, with Mycobacterium bovis BCG serving as the primary immunotherapy, particularly for non-muscle-invasive bladder cancer (NMIBC). However, the mechanisms underlying BCG’s antitumor effects and the potential of non-tuberculous mycobacteria like Mycobacterium brumae remain unclear. This study investigates the antitumor effects of M. bovis BCG and M. brumae on BC cell migration, invasion, and anchorage-independent growth. BC cell lines representing different stages of tumor differentiation were treated with either M. bovis BCG or M. brumae. Cell migration was assessed through wound healing and transwell assays, invasiveness by transwell invasion assays, MMP-9 production by gelatin zymography, and anchorage-independent growth via soft agar colony formation. Both mycobacteria inhibited individual cell migration across all BC lines, while collective migration was only reduced in intermediate-grade cells. Both treatments also reduced invasiveness, associated with decreased MMP-9 production. Furthermore, M. brumae inhibited anchorage-independent growth across all BC lines, while M. bovis BCG had a more selective effect, primarily inhibiting growth in high-grade cells. In conclusion, both mycobacteria reduce migration, invasion, and anchorage-independent growth of BC cells, with their effectiveness varying by species and tumor differentiation grade. Keywords: BCG; non-muscle invasive; bacteriotherapy Author: Marc Bach-Griera, Alba Hernández and Esther Julián Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Tue Dec 3 12:38:02 2024 CET ModDate: Tue Dec 3 12:41:24 2024 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 9051862 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AIQCCL+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 CAEOCO+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 WHPZNE+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 SNLFUF+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 MOCBVF+VnURWPalladioL Type 1 Custom yes yes yes 32 0 LEVXPH+EURM10 Type 1 Builtin yes yes yes 37 0 DYUFJK+CMSY10 Type 1 Builtin yes yes yes 70 0 CHIAOF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 82 0 CHIAPG+PalatinoLinotype CID TrueType Identity-H yes yes yes 85 0 CHIAPH+PalatinoLinotype TrueType WinAnsi yes yes no 91 0 CHIAPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 94 0 CHICCD+PalatinoLinotype TrueType MacRoman yes yes no 97 0 CHIEPD+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 100 0 CHIAOF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 116 0 CHIAPG+PalatinoLinotype CID TrueType Identity-H yes yes yes 119 0 CHIAPH+PalatinoLinotype TrueType WinAnsi yes yes no 125 0 CHIAPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 128 0 CHICCD+PalatinoLinotype TrueType MacRoman yes yes no 131 0 CHIAOF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 146 0 CHIAPG+PalatinoLinotype CID TrueType Identity-H yes yes yes 149 0 CHIAPH+PalatinoLinotype TrueType WinAnsi yes yes no 155 0 CHIAPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 158 0 CHICCD+PalatinoLinotype TrueType MacRoman yes yes no 161 0 CHIEPD+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 164 0 CHIAOF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 176 0 CHIAPG+PalatinoLinotype CID TrueType Identity-H yes yes yes 179 0 CHIAPH+PalatinoLinotype TrueType WinAnsi yes yes no 185 0 CHIAPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 188 0 CHICCD+PalatinoLinotype TrueType MacRoman yes yes no 191 0 CHIEPD+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 194 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-02-14 02:54:04 CET RepresentationInformation: ijms-25-12997.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-02-12 07:37:52 CET Size: 9051862 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 420 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: BCG and M. brumae Infections Decrease Collective Migration Capability in Intermediate-Grade BC Cells Destination: subsection.2.1 Item: Title: Mycobacteria Infection Decreases the Single Cell Migration Capability of BC Cells Destination: subsection.2.2 Item: Title: M. brumae Treatment Alone Decreases Anchorage-Independent Growth in BC Cell Lines Destination: subsection.2.3 Item: Title: M. brumae and BCG Treatments Decrease Cell Invasion by Inhibiting Active MMP-9 Secretion Destination: subsection.2.4 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Bacterial Strains and Infection Procedure Destination: subsection.4.1 Item: Title: Cell Culture Conditions Destination: subsection.4.2 Item: Title: Collective Cell Migration Through Wound-Healing Assay Destination: subsection.4.3 Item: Title: Single Cell Migration and Invasion Through Transwells Assays Destination: subsection.4.4 Item: Title: Anchorage Independent Growth by Soft Agar Colony Formation Assay Destination: subsection.4.5 Item: Title: Secretion of Matrix Metalloproteinases (MMPs) by Zymography Destination: subsection.4.6 Item: Title: Statistical Analyses Destination: subsection.4.7 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Mycobacteria Treatment Inhibits Bladder Cancer Cell Migration, Invasion, and Anchorage-Independent Growth Author: Marc Bach-Griera, Alba Hernández and Esther Julián Subject: Bladder cancer (BC) is a highly recurrent and invasive malignancy, with Mycobacterium bovis BCG serving as the primary immunotherapy, particularly for non-muscle-invasive bladder cancer (NMIBC). However, the mechanisms underlying BCG’s antitumor effects and the potential of non-tuberculous mycobacteria like Mycobacterium brumae remain unclear. This study investigates the antitumor effects of M. bovis BCG and M. brumae on BC cell migration, invasion, and anchorage-independent growth. BC cell lines representing different stages of tumor differentiation were treated with either M. bovis BCG or M. brumae. Cell migration was assessed through wound healing and transwell assays, invasiveness by transwell invasion assays, MMP-9 production by gelatin zymography, and anchorage-independent growth via soft agar colony formation. Both mycobacteria inhibited individual cell migration across all BC lines, while collective migration was only reduced in intermediate-grade cells. Both treatments also reduced invasiveness, associated with decreased MMP-9 production. Furthermore, M. brumae inhibited anchorage-independent growth across all BC lines, while M. bovis BCG had a more selective effect, primarily inhibiting growth in high-grade cells. In conclusion, both mycobacteria reduce migration, invasion, and anchorage-independent growth of BC cells, with their effectiveness varying by species and tumor differentiation grade. 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