Establishment and characterization of a multi-drug resistant cell line for canine mammary tumors

Background and purposeCanine mammary tumors are the most common tumor disease of female dogs, and adjuvant chemotherapy often results in multi-drug resistance. Currently, the mechanisms underlying the development of tumor multi-drug resistance are unclear. The translation of research applications that can be used to effectively overcome tumor resistance is similarly hampered. Therefore, it is urgent to construct multi-drug resistance models of canine mammary tumors that can be used for research, to explore the mechanisms and means of overcoming resistance.Materials and methodsIn this study, the canine triple negative breast cancer cell line CMT-7364 was induced to develop multidrug resistance using doxorubicin by high-dose drug pulse method. The drug resistance and the expression of drug transport pumps of the cells was verified by CCK8 assay, immunoblotting, qPCR and immunofluorescence. Next, we used scratch assay and Transwell invasion assay to compare the migration and invasion abilities of the two cell lines and examined the expression of EMT-related proteins in both using immunoblotting. The differences of transcriptome between parental and drug-resistant cell lines were detected by RNA-seq sequencing. Finally, mouse xenograft models of drug-resistant and parental cell lines were constructed to evaluate the tumorigenic ability.ResultsAfter more than 50 generations of continuous passages stimulated by high-dose drug pulse method, the morphology of drug-resistant cell line CMT-7364/R tended to be mesenchymal-like and heterogeneous under light microscopy compared with the parental cell line CMT-7364/S, and developed resistance to doxorubicin and other commonly used chemotherapeutic drugs. In CMT-7364/R, BCRP was expressed at higher levels at both transcriptional and protein levels, while P-glycoprotein was not significantly different. Secondly, the migration and invasion ability of CMT-7364/R was significantly enhanced, with decreased expression of E-cadherin and increased expression of vimentin and mucin 1-N terminus. Finally, mouse xenograft models were constructed, while there was no significant difference in the volume of masses formed at 21 days.ConclusionIn summary, by using the canine mammary tumor cell line CMT-7364/S as the parental cell line, we successfully constructed a multidrug-resistant CMT-7364/R with high-dose drug pulse methods. Compared to its parental cell line, CMT-7364/R has decreased growth rate, overexpression of BCRP and increased migration and invasion ability due to EMT. The results of this study showed that CMT-7364/R might serve as a model for future studies on tumor drug resistance

First Observations of Mars Atmosphere and Ionosphere with Tianwen-1 Radio-Occultation Technique on 5 August 2021

The radio-occultation technique can provide vertical profiles of planetary ionospheric and atmospheric parameters, which merit the planetary-climate and space-weather scientific research so far. The Tianwen-1 one-way single-frequency radio-occultation technique was developed to retrieve Mars ionospheric and atmospheric parameters. The first radio-occultation event observation experiment was conducted on 5 August 2021. The retrieved excess Doppler frequency, bending angle, refractivity, electron density, neutral mass density, pressure and temperature profiles are presented. The Mars ionosphere M1 (M2) layer peak height is at 140 km (105 km) with a peak density of about 3.7 × 1010 el/m3 (5.3 × 1010 el/m3) in the retrieved electron-density profile. A planetary boundary layer (−2.35 km~5 km), a troposphere (temperature decreases with height) and a stratosphere (24 km–40 km) clearly appear in the retrieved temperature profile below 50 km. Results show that Tianwen-1 radio occultation data are scientifically reliable and useful for further Mars climate and space-weather studies

First Observations of Mars Atmosphere and Ionosphere with Tianwen-1 Radio-Occultation Technique on 5 August 2021

The radio-occultation technique can provide vertical profiles of planetary ionospheric and atmospheric parameters, which merit the planetary-climate and space-weather scientific research so far. The Tianwen-1 one-way single-frequency radio-occultation technique was developed to retrieve Mars ionospheric and atmospheric parameters. The first radio-occultation event observation experiment was conducted on 5 August 2021. The retrieved excess Doppler frequency, bending angle, refractivity, electron density, neutral mass density, pressure and temperature profiles are presented. The Mars ionosphere M1 (M2) layer peak height is at 140 km (105 km) with a peak density of about 3.7 × 1010 el/m3 (5.3 × 1010 el/m3) in the retrieved electron-density profile. A planetary boundary layer (−2.35 km~5 km), a troposphere (temperature decreases with height) and a stratosphere (24 km–40 km) clearly appear in the retrieved temperature profile below 50 km. Results show that Tianwen-1 radio occultation data are scientifically reliable and useful for further Mars climate and space-weather studies