Bijie is located at a typical karst landform of Southwestern Guizhou, which presented high geological background values of potentially toxic elements (PTEs). Recently, whether PTE of wheat in Bijie is harmful to human health has aroused people’s concern. To this end, the objectives of this study are to determine the concentrations of PTE [chromium (Cr), nickel (Ni), arsenic (As), lead (Pb), cadmium (Cd), and fluorine (F)] in wheat grains, identify contaminant sources, and evaluate the probabilistic risks to human beings. A total of 149 wheat grain samples collected from Bijie in Guizhou were determined using the inductively coupled plasma mass spectrometer (ICP-MS) and fluoride-ion electrode methods. The mean concentrations of Cr, Ni, As, Cd, Pb, and F were 3.250, 0.684, 0.055, 0.149, 0.039, and 4.539 mg/kg, respectively. All investigated PTEs met the standard limits established by the Food and Agriculture Organization except for Cr. For the source identification, Cr and Pb should be originated from industry activities, while Ni, As, and Cd might come from mixed sources, and F was possibly put down to the high geological background value. The non-carcinogenic and carcinogenic health risks were evaluated by the probabilistic approach (Monte Carlo simulation). The mean hazard quotient (HQ) values in the three populations were lower than the safety limit (1.0) with the exception of As (children: 1.03E+00). However, the mean hazard index (HI) values were all higher than 1.0 and followed the order: children (2.57E+00) > adult females (1.29E+00) > adult males (1.12E+00). In addition, the mean carcinogenic risk (CR) values for Cr, As, Pb, and Cd in three populations were all higher than 1E-06, which cannot be negligible. The mean threshold CR (TCR) values were decreased in the order of children (1.32E-02) > adult females (6.61E-03) > adult males (5.81E-03), respectively, all at unacceptable risk levels. Moreover, sensitivity analysis identified concentration factor (CW) as the most crucial parameter that affects human health. These findings highlight that co-exposure of PTE in wheat grains revealed a probabilistic human health risk. Corresponding measures should be undertaken for controlling pollution sources and reducing the risks for the local populace.