Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 Reporter
Transcription factors Sox2 and Oct4 are essential in maintaining the pluripotency of embryonic stem cells and conferring stemness in cancer stem-like (CSL) cells. SORE6, an in-vitro reporter system, was designed to quantify the transcription activity of Sox2/Oct4 and identify CSL cells in non-hemato...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-07-01
|
| Series: | Current Issues in Molecular Biology |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1467-3045/43/2/41 |
| _version_ | 1797519748215341056 |
|---|---|
| author | Jing Li Moinul Haque Chuquan Shang Bardes Hassan Dongzhe Liu Will Chen Raymond Lai |
| author_facet | Jing Li Moinul Haque Chuquan Shang Bardes Hassan Dongzhe Liu Will Chen Raymond Lai |
| author_sort | Jing Li |
| collection | DOAJ |
| description | Transcription factors Sox2 and Oct4 are essential in maintaining the pluripotency of embryonic stem cells and conferring stemness in cancer stem-like (CSL) cells. SORE6, an in-vitro reporter system, was designed to quantify the transcription activity of Sox2/Oct4 and identify CSL cells in non-hematologic cancers. Using SORE6, we identified and enriched CSL cells in ALK-positive anaplastic large cell lymphoma (ALK + ALCL). Two ALK + ALCL cell lines, SupM2 and UCONN-L2, contained approximately 20% of SORE6+ cells, which were purified based on their expression of green fluorescent protein. We then performed functional studies using single-cell clones derived from SORE6− and SORE6+ cells. Compared to SORE6− cells, SORE6+ cells were significantly more chemoresistant and clonogenic in colony-formation assays. Sox2/Oct4 are directly involved in conferring these CSL properties, since the shRNA knockdown of Sox2 in SORE6+ significantly lowered their chemoresistance, while enforced expression of Sox2/Oct4 in SORE6− cells produced opposite effects. Using Western blots, we found that the expression and subcellular localization of Sox2/Oct4 were similar between SORE6− and SORE6+ cells. However, in SORE6+ but not SORE6− cells, Sox2 and Oct4 abundantly bound to a probe containing the SORE6 consensus sequence. c-Myc, previously shown to regulate cancer stemness in ALK + ALCL, regulated the SORE6 activity. In conclusion, SORE6 is useful in identifying/enriching CSL cells in ALK + ALCL. |
| first_indexed | 2024-03-10T07:47:11Z |
| format | Article |
| id | doaj.art-2e4f33458aed4657b790a84f2169e046 |
| institution | Directory Open Access Journal |
| issn | 1467-3037 1467-3045 |
| language | English |
| last_indexed | 2024-03-10T07:47:11Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Current Issues in Molecular Biology |
| spelling | doaj.art-2e4f33458aed4657b790a84f2169e0462023-11-22T12:30:51ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452021-07-0143254355710.3390/cimb43020041Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 ReporterJing Li0Moinul Haque1Chuquan Shang2Bardes Hassan3Dongzhe Liu4Will Chen5Raymond Lai6Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, CanadaDepartment of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, CanadaDepartment of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, CanadaDepartment of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, CanadaDepartment of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, CanadaDepartment of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, CanadaDepartment of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, CanadaTranscription factors Sox2 and Oct4 are essential in maintaining the pluripotency of embryonic stem cells and conferring stemness in cancer stem-like (CSL) cells. SORE6, an in-vitro reporter system, was designed to quantify the transcription activity of Sox2/Oct4 and identify CSL cells in non-hematologic cancers. Using SORE6, we identified and enriched CSL cells in ALK-positive anaplastic large cell lymphoma (ALK + ALCL). Two ALK + ALCL cell lines, SupM2 and UCONN-L2, contained approximately 20% of SORE6+ cells, which were purified based on their expression of green fluorescent protein. We then performed functional studies using single-cell clones derived from SORE6− and SORE6+ cells. Compared to SORE6− cells, SORE6+ cells were significantly more chemoresistant and clonogenic in colony-formation assays. Sox2/Oct4 are directly involved in conferring these CSL properties, since the shRNA knockdown of Sox2 in SORE6+ significantly lowered their chemoresistance, while enforced expression of Sox2/Oct4 in SORE6− cells produced opposite effects. Using Western blots, we found that the expression and subcellular localization of Sox2/Oct4 were similar between SORE6− and SORE6+ cells. However, in SORE6+ but not SORE6− cells, Sox2 and Oct4 abundantly bound to a probe containing the SORE6 consensus sequence. c-Myc, previously shown to regulate cancer stemness in ALK + ALCL, regulated the SORE6 activity. In conclusion, SORE6 is useful in identifying/enriching CSL cells in ALK + ALCL.https://www.mdpi.com/1467-3045/43/2/41anaplastic large cell lymphomaSORE6cancer stem-like cellsSox2Oct4 |
| spellingShingle | Jing Li Moinul Haque Chuquan Shang Bardes Hassan Dongzhe Liu Will Chen Raymond Lai Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 Reporter Current Issues in Molecular Biology anaplastic large cell lymphoma SORE6 cancer stem-like cells Sox2 Oct4 |
| title | Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 Reporter |
| title_full | Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 Reporter |
| title_fullStr | Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 Reporter |
| title_full_unstemmed | Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 Reporter |
| title_short | Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using the SORE6 Reporter |
| title_sort | identification and characterization of cancer stem like cells in alk positive anaplastic large cell lymphoma using the sore6 reporter |
| topic | anaplastic large cell lymphoma SORE6 cancer stem-like cells Sox2 Oct4 |
| url | https://www.mdpi.com/1467-3045/43/2/41 |
| work_keys_str_mv | AT jingli identificationandcharacterizationofcancerstemlikecellsinalkpositiveanaplasticlargecelllymphomausingthesore6reporter AT moinulhaque identificationandcharacterizationofcancerstemlikecellsinalkpositiveanaplasticlargecelllymphomausingthesore6reporter AT chuquanshang identificationandcharacterizationofcancerstemlikecellsinalkpositiveanaplasticlargecelllymphomausingthesore6reporter AT bardeshassan identificationandcharacterizationofcancerstemlikecellsinalkpositiveanaplasticlargecelllymphomausingthesore6reporter AT dongzheliu identificationandcharacterizationofcancerstemlikecellsinalkpositiveanaplasticlargecelllymphomausingthesore6reporter AT willchen identificationandcharacterizationofcancerstemlikecellsinalkpositiveanaplasticlargecelllymphomausingthesore6reporter AT raymondlai identificationandcharacterizationofcancerstemlikecellsinalkpositiveanaplasticlargecelllymphomausingthesore6reporter |